4-Mercaptopyrrolidine derivatives as farnesyl transferase inhibitors

ABSTRACT

Pharmaceutical compositions comprising an inhibitor of ras farnesylation of formula (I) wherein, R 1  is for example H and further values as defined in the specification; R 2  is for example H and further values as defined in the specification; R 3  is for example H or a substituent having values as defined in the specification; p is 0-3 in which R 3  values can be the same or different; L is a linking moiety for example —CO—NH 2 — and further values as defined in the specification; A is selected from phenyl; naphthyl; a 5-10 membered monocyclic or bicyclic heteroaryl ring containing up to 5 heteroatoms where the heteroatoms are independently selected from O, N and S; or a —S—S— dimer thereof when R 2 =H; or an enantiomer, diastereoisomer, pharmaceutically acceptable salt, prodrug or solvate thereof together with a pharmaceutically acceptable diluent or carrier. A particular use is cancer therapy.

This invention relates to compounds that inhibit farnesylation of mutantras gene products through inhibition of the enzyme farnesyl-proteintransferase (FPTase). The invention also relates to methods ofmanufacturing the compounds, pharmaceutical compositions and methods oftreating diseases, especially cancer, which are mediated throughfarnesylation of ras.

Cancer is believed to involve alteration in expression or function ofgenes controlling cell growth and differentiation. Whilst not wishing tobe bound by theoretical considerations the following text sets out thescientific background to ras in cancer. Ras genes are frequently mutatedin tumours. Ras genes encode guanosine triphosphate (GTP) bindingproteins which are believed to be involved in signal transduction,proliferation and malignant transformation, H-, K- and N-ras genes havebeen identified as mutant forms of ras (Barbacid M. Ann. Rev. Biochem.1987, 56: 779-827). Post translational modification of ras protein isrequired for biological activity. Farnesylation of ras catalysed byFPTase is believed to be an essential step in ras processing. It occursby transfer of the farnesyl group of farnesyl pyrophosphate (FPP) to acysteine at the C-terminal tetrapeptide of ras in a structural motifcalled the CAAX box. After further post-translational modifications,including proteolytic cleavage at the cysteine residue of the CAAX boxand methylation of the cysteine carboxyl, ras is able to attach to thecell membrane for relay of growth signals to the cell interior. Innormal cells activated ras is believed to act in conjunction with growthfactors to stimulate cell growth. In tumour cells it is believed thatmutations in ras cause it to stimulate cell division even in the absenceof growth factors (Travis J. Science 1993, 260: 1877-1878), possiblythrough being permanently in GTP activated form rather than cycled backto GDP inactivated form. Inhibition of farnesylation of mutant ras geneproducts will stop or reduce activation.

One class of known inhibitors of farnesyl transferase is based onfarnesyl pyrophosphate analogues: see for example European patentapplication EP 534546 from Merck. Inhibitors of farnesyl transferasebased on mimicry of the CAAX box have been reported. Reiss (1990) inCell 62, 81-8 disclosed tetrapeptides such as CVIM (Cys-Val-Ile-Met).James (1993) in Science 260, 1937-1942 disclosed benzodiazepine basedpeptidomimetic compounds. After earliest priority date of the presentinvention Lerner (1995) in J. Biol. Chem. 270, 26802 and Eisai inInternational Patent Application WO 95/25086 disclosed furtherpeptidomimetic compounds based on Cys as the first residue. Also afterthe earliest priority date of the present invention Bristol-Myers Squibbin European Patent Application EP 696593 disclosed for the first timefarnesyl transferase inhibitors having a 4-sulfanylpyrrolidine residuein the first position.

According to one aspect of the present invention there is provided apharmaceutical composition comprising an inhibitor of ras farnesylationof Formula I:

wherein:

R¹ is selected from H; —C₁₋₄alkyl; —C₁₋₃alkylene-Ph optionally mono ordi-substituted on Ph with substituents selected from C₁₋₄alkyl, halogen,OH, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, amino, C₁₋₄alkylamino,di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, nitro, cyano, carboxy, carbamoyl,C₁₋₄alkoxycarbonyl, thiol, C₁₋₄alkylsulfanyl,C₁₋₄alkylsulfinyl,C₁₋₄alkylsulfonyl and sulfonamido; —CO—C₁₋₄alkyl;—CO—O—C₁₋₄alkyl; —CO—O—C₂₋₄alkenyl; —CO—O—(CH₂)_(n)Ph optionallysubstituted on Ph as defined for substitution on Ph inR¹=—C₁₋₃alkylene-Ph above and n=0-4; —C₁₋₄alkylene-CONR⁴R⁵ where R⁴ & R⁵are independently selected from H and C₁₋₄alkyl; and —C₁₋₄alkylene-COOR⁶where R⁶ is selected from H, C₁₋₄alkyl;

R² is selected from H; —C₁₋₄alkyl; —C₁₋₃alkylene-Ph optionallysubstituted on Ph as defined for substitution on Ph inR¹=—C₁₋₃alkylene-Ph above; —COC₁₋₄alkyl; and —COOC₁₋₄alkyl;

R³ is selected from H; OH; CN; CF₃; NO₂; —C₁₋₄alkyl; —C₁₋₄alkylene-R⁷where R⁷ is selected from phenyl, naphthyl, a 5-10 membered monocyclicor bicyclic heteroaryl ring containing upto 5 heteroatoms selected fromO,N and S and any aryl ring in R⁷ is optionally substituted as definedfor substitution on the Ph group in R¹=—C₁₋₃alkylene-Ph above; R⁷;C₂₋₄alkenyl; halogen; —(CH₂)_(n)COOR⁸ where n=0-3 and R⁸ represents H,C₁₋₄alkyl, or C₂₋₄alkenyl; —CONR⁹R¹⁰ where R⁹ and R¹⁰ independentlyrepresent H, C₁₋₄alkyl, C₂₋₄alkenyl, —O—C₁₋₄alkyl, —O—C₂₋₄alkenyl,—C₁₋₃alkylenePh optionally substituted as defined for this group for R¹above; —CON(R¹¹)OR¹² where R¹¹ and R¹² independently represent H,C₁₋₄alkyl and C₂₋₄alkenyl; a group of Formula II, —CONR¹³—CHR¹⁴—COOR¹⁷,where R¹³ is H or C₁₋₄alkyl, R¹⁷ is H or C₁₋₆alkyl, R¹⁴ is selected fromthe side chain of a lipophilic amino acid. carbamoylC₁₋₄alkyl,N-(monoC₁₋₄alkyl)carbamoylC₁₋₄alkyl andN-(diC₁₋₄alkyl)carbamoylC₁₋₄alkyl, the group of Formula II having L or Dconfiguration at the chiral alpha carbon in the corresponding free aminoacid; a lactone of formula:

 C₁₋₄alkyl monosubstituted on carbon with ═N—OH; a group of Formula—X—R¹⁵ where X is selected from O, CO, CH₂, S, SO, SO₂ and R¹⁵ isselected from C₁₋₆alkyl, phenyl, naphthyl, a 5-10 membered monocyclic orbicyclic heteroaryl ring containing upto 5 heteroatoms selected from O,Nand S and any aryl ring in R¹⁵ is optionally substituted as defined forthe Ph group in R¹=—C₁₋₃alkylene-Ph;

p is 0-3 in which R³ values can be the same or different;

L is a linking moiety selected from the following groups written fromleft to right in Formula I: —CO—NR¹⁶— where R¹⁶ is selected from H,C₁₋₄alkyl, C₁₋₄alkylene-Z, —CO—C₁₋₄alkylene-Z, —CO—C₁₋₆alkyl, —COZ, Zand Z is selected from —O—C₁₋₄alkyl, phenyl, naphthyl, a 5-10 memberedmonocyclic or bicyclic heteroaryl ring containing upto 5 heteroatomsselected from O, N and S and any aryl ring in R¹⁶ is optionallysubstituted as defined for the Ph group in R¹=—C₁₋₃alkylene-Ph;—CH₂—NR¹⁸— where R¹⁸ represents any value defined for R¹⁶; —CH₂S—;—CH₂O—; —CH₂—CHR¹⁹— where R¹⁹ represents any value defined for R¹⁶;—CH═CR²⁰— where R²⁰ represents any value defined for R¹⁶; —CH₂NR²¹—T—where R²¹ represents any value defined for R¹⁶, T represents —(CH₂)_(n)—where n is 1-4 and T is optionally monosubstituted with R²² where R²²represents any value for R¹⁶ other than H; —CH₂NR²³—SO₂— where R²³represents any value defined for R¹⁶; —CH₂₋—NR²⁴—CO—T— where R²⁴represents any value defined for R¹⁶, T represents —(CH₂)_(n)— where nis 0-4 and T is optionally monosubstituted with R²⁹ where R²⁹ representsany value for R¹⁶ other than H; —CO—NR²⁵—T— where R²⁵ represents anyvalue defined for R¹⁶, T represents —(CH₂)_(n)— where n is 1-4 and T isoptionally monosubstituted with R²⁶ where R²⁶ represents any value forR¹⁶ other than H; —CH₂S—T— where T represents —(CH₂)_(n)— where n is 1-4and T is optionally monosubstituted with R²⁷ where R²⁷ represents anyvalue for R¹⁶ other than H; —CH₂O—T— where T represents —(CH₂)_(n)—where n is 1-4 and T is optionally monosubstituted with R²⁸ where R²⁸represents any value for R¹⁶ other than H;

A is selected from phenyl; naphthyl; a 5-10 membered monocyclic orbicyclic heteroaryl ring containing upto 5 heteroatoms where theheteroatoms are independently selected from O, N & S; or a —S—S— dimerthereof when R²═H; or a N-oxide thereof; or an enantiomer,diastereoisomer, pharmaceutically acceptable salt, prodrug or solvatethereof together with a pharmaceutically acceptable diluent or carrier.

Preferably R¹ is selected from H; —CO—O—(CH₂)_(n)Ph optionallysubstituted on Ph as defined for R¹=—C₁₋₃alkylene-Ph and n=0-4;—CO—O—C₂₋₄alkenyl; —CO—C₁₋₄alkyl; —C₁₋₄alkylene-CONR⁴R⁵ where R⁴ & R⁵are independently selected from H, C₁₋₄alkyl,

Preferably R² is selected from H and —CO—C₁₋₄alkyl.

Preferably L is selected from —CH₂—NR¹⁸—; —CH₂NR²¹—T.

Preferably A is selected from phenyl, naphthyl, pyridyl and thienyl.

Preferably combinations of R³ and p are selected from:

i) R³ is selected from a group of Formula II; —C₁₋₄-alkylR⁷; —O—R⁷ and;R⁷; and p=1-3 with the proviso that one value of R³ is a group ofFormula II;

ii) p=0 with the proviso that A is naphthyl and L is —CH₂NR²¹—T;

iii) p=1 with the proviso that R³=a group of Formula II and A isnaphthyl.

In another embodiment of the invention it is preferred that:

R¹ is selected from H; —C₁₋₄alkyl, —C₁₋₃alkylene-Ph optionally mono ordi-substituted on Ph with substituents selected from C₁₋₄alkyl, halogen,OH, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, amino, C₁₋₄alkylamino,di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, thiol, C₁₋₄alkylthio, nitro,cyano, carboxy, carbamoyl, C₁₋₄alkoxycarbonyl, C₁₋₄alkylsulfinyl,C₁₋₄alkylsulfonyl, sulfonamido; —CO—C₁₋₄alkyl; —CO—O—C₁₋₄alkyl;—CO—O—C₂₋₄alkenyl; —CO—O—CH₂₋—Ph optionally mono- or di-substituted onphenyl with substituents selected from C₁₋₄alkyl, halogen, OH,C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, amino, C₁₋₄alkylamino,di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, thiol, C₁₋₄alkylthio, nitro,cyano, carboxy, carbamoyl, C₁₋₄alkoxycarbonyl, C₁₋₄ alkylthiono,C₁₋₄alkylsulfonyl, sulfonamido; —C₁₋₄alkylene-CONR⁴R⁵ where R⁴ & R⁵ areindependently selected from H, C₁₋₄alkyl; —C₁₋₄alkylene-COOR⁶ where R⁶is selected from H, C₁₋₄alkyl;

R² is selected from H; —C₁₋₄alkyl; —C₁₋₃alkylene-Ph; —COC₁₋₄alkyl;—COOC₁₋₄alkyl;

R³ is selected from H; OH; CN; CF₃; NO₂; —C₁₋₄ alkyl, —C₁₋₄alkylene-R⁷where R⁷ is selected from phenyl, naphthyl, a 5-10 membered monocyclicor bicyclic heteroaryl ring containing upto 3 heteroatoms selected fromO,N and S; C₂₋₄alkenyl; halogen; —(CH₂)_(n)COOR⁸ where n=0-3 and R⁸represents H, C₁₋₄alkyl, C₂₋₄alkenyl; —CONR⁹R¹⁰ where R⁹ and R¹⁰independently represent H, C₁₋₄alkyl, C₂₋₄alkenyl, —O—C₁₋₄alkyl,—O—C₂₋₄alkenyl; —CON(R¹¹)OR¹² where R¹¹ and R¹² independently representH, C₁₋₄alkyl and C₂₋₄alkenyl; a group of Formula II,—CONR¹³—CHR¹⁴—COOR¹⁷, where R¹³ is H or C₁₋₄alkyl, R¹⁷ is H orC₁₋₆alkyl, R¹⁴ is the side chain of a lipophilic amino acid with L or Dconfiguration at the chiral alpha carbon in the corresponding free aminoacid; C₁₋₄alkyl monosubstituted on carbon with ═N—OH; —SO—C₁₋₄alkyl;—SO₂—C₁₋₄alkyl; a group of Formula —X—R¹⁵ where X is selected from CO,CH₂, S, SO, SO₂ and R¹⁵ is selected from C₁₋₆alkyl, phenyl, naphthyl, a5-10 membered monocyclic or bicyclic heteroaryl ring containing upto 3heteroatoms selected from O,N and S;

p is 0-3 in which R³ values can be the same or different;

L is a linking moiety selected from the following groups written fromleft to right in Formula I: —CO—NR¹⁶— where R¹⁶ is selected from H,C₁₋₄alkyl, C₁₋₄alkylene-Z and Z is selected from —O—C₁₋₄alkyl, phenyl,naphthyl, a 5-10 membered monocyclic or bicyclic heteroaryl ringcontaining upto 3 heteroatoms selected from O, N and S; —CH₂₋—NR¹⁸—where R¹⁸ represents any value defined for R¹⁶; —CH₂S—; —CH₂O—;—CH₂₋CHR¹⁹— where R¹⁹ represents any value defined for R¹⁶; —CH═CR²⁰—where R²⁰ represents any value defined for R¹⁶; —CH₂NR²¹—T— where R²¹represents any value defined for R¹⁶, T represents —(CH₂)_(n)— where nis 1-4 and T is optionally monosubstituted with R²² where R²² representsany value for R¹⁶ other than H, and provided at least one of R²¹ and R²²is H; —CH₂NR²³—SO₂— where R²³ represents any value defined for R¹⁶;—CH₂₋NR²⁴—CO—T— where R²⁴ represents any value defined for R¹⁶, Trepresents —(CH₂)_(n)— where n is 0-4 and T is optionallymonosubstituted with R²⁹ where R²⁹ represents any value for R¹⁶ otherthan H, and provided at least one of R²⁴ and R²⁹ is H; —CO—NR²⁵—T— whereR²⁵ represents any value defined for R¹⁶, T represents —(CH₂)_(n)— wheren is 1-4 and T is optionally monosubstituted with R²⁶ where R²⁶represents any value for R¹⁶ other than H, and provided at least one ofR²⁴ and R²⁵ is H; —CH₂S—T— where T represents —(CH₂)_(n)— where n is 1-4and T is optionally monosubstituted with R²⁷ where R²⁷ represents anyvalue for R¹⁶ other than H; —CH₂O—T— where T represents —(CH₂)_(n)—where n is 1-4 and T is optionally monosubstituted with R²⁸ where R²⁸represents any value for R¹⁶ other than H;

A is selected from phenyl; naphthyl; a 5-10 membered monocyclic orbicyclic heteroaryl ring containing upto 3 or 5 heteroatoms in the caseof monocyclic and bicyclic rings respectively where the heteroatoms areindependently selected from O, N & S; or a —S—S— dimer thereof whenR²=H.

A preferred pharmaceutical composition is in the form of a tablet.

According to another aspect of the invention there is provided acompound of Formula I, II, IV or V for use as a medicament.

According to another aspect of the invention there is provided acompound of Formula I, IIII, IV or V for use in preparation of amedicament for treatment of a disease mediated through farnesylation ofras.

Many compounds of Formula I are a feature of this invention and inparticular according to another aspect of the invention there isprovided a compound of any of the following classes i), ii) or iii):

wherein:

X¹ is selected from H; C₁₋₆alkyl; hydroxyC₁₋₆alkyl, C₁₋₆alkoxyC₁₋₆alkyl,C₁₋₆alkylcarbonyl; hydroxyC₁₋₆alkylcarbonyl;C₁₋₆alkoxyC₁₋₆alkylcarbonyl;

A is selected from phenyl, naphthyl or a 5-10 membered heterocyclic ringhaving upto 5 heteroatoms selected from O, N and S:

X² is selected from H; phenyl; phenylC₁₋₆alkyl; a 5-6 memberedheteroaryl ring containing upto 3 heteroatoms selected from O, N and Soptionally linked to A by C₁₋₆alkyl; and X² is optionally substituted onany ring as defined for phenyl in R¹=—C₁₋₃alkylene-Ph in claim 1;

X³ is selected from H; C₁₋₆alkyl;

X⁴ is selected from C₁₋₆alkylsulfanyl; C₁₋₆alkylsulfinyl;C₁₋₆alkylsulfonyl; carbamoyl; N-(C₁₋₆alkyl)carbamoyl;N-(diC₁₋₆alkyl)carbamoyl; and hydroxy or a C₁₋₄alkyl ether thereof:

 wherein:

X⁵ is selected from —CO—C₁₋₄alkyl-Ph; —CO—C₁₋₆alkyl;—CO—C₁₋₄alkyl-heteroaryl where heteroaryl is a 5-10 membered heteroarylring containing upto 5 heteroatoms selected from O, N and S and Ph orheteroaryl are optionally substituted as defined for Ph inR¹=—C₁₋₃alkylene-Ph; C₁₋₄alkyloxyC₁₋₄alkyl;

A is naphthyl or a 10 membered heterocyclic ring having upto 5heteroatoms selected from O, N and S; R³ and p are as defined in claim1;

 wherein:

X⁶ has any value defined for X⁵ in ii) above;

X⁷ is Ph optionally substituted as defined for Ph inR¹=—C₁₋₃alkylene-Ph;

A is Ph or naphthyl or a 5-10 membered heterocyclic ring having upto 5heteroatoms selected from O, N and S;

R³ and p are as defined above: or a N-oxide, pharmaceutically acceptablesalt, prodrug or solvate thereof.

Preferred values for compounds of class i) include,

X¹ is selected from H and C₁₋₆alkoxyC₁₋₆alkyl;

X² is selected from H; phenyl or phenylC₁₋₆alkyl;

X⁴ is C₁₋₆alkylsulfanyl;

A is selected from phenyl or naphthyl; Other preferred values for X⁴ are—OMe and the lactone which can be formed when X⁴ is OH and X³ is H.

Preferred values for compounds of class ii) include p is 0.

Preferred values for compounds of class iii) include,

X⁷ is Ph;

A is Ph;

p is 0.

In another embodiment of the invention there is provided a compound ofFormula I in which: R¹ is selected from H or C₁₋₄alkyl; R² is selectedfrom H, C₁₋₄alkyl, —COC₁₋₄alkyl, —C₁₋₄alkylPh; L is selected from thefollowing values as defined herein. CONR¹⁶, CH₂S, CH₂O, CH₂CHR¹⁹,CH═CHR²⁰, CH₂NR²⁴COT, CONR²⁵T, CH₂ST and CH₂OT; and values for A, R³ andp are as defined herein, with the proviso that2-(benzylcarbamoyl)-4-sulfanylpyrollidine and4-(acetylsulfonyl)-2(benzylcarbamoyl)-pyrrolidine are excluded. It isbelieved that the excluded compounds were disclosed as intermediates forbeta-lactam antibiotic synthesis in Japanese patent application 60233076(Sumitomo Chemical).

According to another aspect of the present invention there is providedany one of the following individual compounds or a pharmaceuticallyacceptable salt thereof:

(2S)-2-{2-Benzyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid methyl ester;

(2S)-2-{2-Benzyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)amino]-benzoylamino}-4-methylsulfanylbutyricacid;

(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester;

(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid;

(2S)-2-({3-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester;

(2S)-2-({3-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl}-amino)-4methylsulfanylbutyricacid;

(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester;

(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid;

(2S,4S)-2-[{N-(4-methoxybenzyl)-N-(naphthalen-1-ylmethyl)-amino}-methyl]-pyrrolidine-4-thiol;

N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-pentanamide;

N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-2-(pyridin-3-yl)-acetamide;

N-((2S,4S)-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-N-(2-naphthalen-1-yl-ethyl)butyramide;

N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-N-(2-naphthalen-1-yl-ethyl)-2-pyridin-3-yl-acetamide;

(2S,4S)-2-{[(3-Methoxypropyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol;

N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-2-(4-methoxy-phenyl)-N-(2-naphthalen-2-yl-ethyl)-acetamide;

(2S,4S)-2-{[(2-(4-Methoxyphenyl)ethyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol;

N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-butyramide;

N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-2-yl-ethyl)-butyramide;

N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-butyramide;

(2S)-2-{3-[([2S,4S]-4-Sulfanyl-pyrrolidin-2-ylmethyl)-(3-methoxy-propyl)-amino]-benzoylamino}-4-methylsulfanyl-butyricacid;

N-([2S,4S]-4Sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-1-yl-ethyl)-butyramide;

(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyricacid; and

(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)butyricacid methyl ester.

According to another aspect of the invention there is provided apharmaceutical composition comprising a compound as defined in any oneFormulas III, IV or V or an individual compound listed above togetherwith a pharmaceutically acceptable diluent or carrier.

According to another aspect of the invention there is provided a processfor preparing compounds of classes i), ii) or iii) as defined abovewhich comprises deprotecting a compound of Formula VI:

wherein X⁸ represents the right hand side of compound classes i), ii) oriii) as defined above, Pr¹ is H or an amino protecting group, Pr² is Hor a thio protecting group and any functional groups in X⁸ areoptionally protected with the proviso that there is at least oneprotecting group and optionally, if desired, converting the product thusobtained into a pharmaceutically acceptable salt thereof.

In an embodiment of the invention:

Examples of values for R¹ include methyl; —CH₂—Ph; —CH₂—Ph substitutedon Ph with nitro, especially 4-nitro: acetyl: BOC; allyloxycarbonyl;—CO—O—CH₂—Ph substituted on Ph with nitro, especially 4-nitro;—CH₂CONH₂.

Examples of values for R² include —COMe and —COOtertbutyl.

Examples of values for R³ include Cl; —COOH; —CONH₂; —SOMe and; —SO₂Me.

When R³ represents —(CH₂)_(n)—COOR⁸ a suitable value for n is 0.

Examples of lipophilic amino acids which contribute their side chain(denoted R¹⁴ within the definition of values for R³) include methionine,phenylglycine, phenylalanine, serine, leucine, isoleucine or valine, Lconfiguration in the corresponding free amino acid is preferred.Examples of amino acid side chains are set out below. A preferred valuefor R¹⁴ is —CH₂—CH₂—S—CH₃. Further preferred values for R¹⁴ are —CH₂—OMeand —CH₂—CH₂—OMe.

When R¹⁷ is H to give a COOH group in Formula II, and R¹⁴ is —CH₂—CH₂—OHthen a lactone can be formed where R¹⁷ and R¹⁴ together form part of adihydrofuran-2-one heterocyclic ring. The same lactone can be formed forcompounds of Formula III where X⁴ is OH and X³ is H.

Amino Acid Side Chain methionine —CH₂—CH₂—S—CH₃ phenylglycine Phphenylalanine —CH₂—Ph serine —CH₂OH or a C₁₋₄alkyl (preferably methyl)ether thereof. leucine —CH₂—CHMe₂ homoserine —CH₂—CH₂—OH or a C₁₋₄alkyl(preferably methyl) ether thereof.

A preferred value for p is 2.

When L is —CH₂NR²¹—T— a suitable value for n is 1. When L is—CH₂—NR²⁴—CO—T— a suitable value for n is 1. When L is —CH₂—NR²⁵—T— asuitable value for n is 1. When L is —CH₂—S—T— a suitable value for nis 1. When L is —CH₂—O—T— a suitable value for n is 1. L is especially—CONH—, —CH₂—NH—, —CH₂NHSO₂—, —CH₂NHCO—.

Examples of values for A when A is heteroaryl are thienyl, pyridyl,quinolyl & quinoxalinyl.

Further preferred values are set out below.

For R¹; 4-nitro-benzyloxycarbonyl; allyloxycarbonyl; carbamoylmethyl;acetyl; phenoxycarbonyl; H.

For R²: Acetylsulfanyl; H.

For R³: Methoxycarbonyl; N-methyl-N-methoxy-carbamoyl; nitro;allyloxycarbonyl; N-methyl-allyloxycarbamoyl; ethoxycarbonyl;3,4-dichloro-benzyl-carbamoyl; hydroxy; carboxy;(2S),4-methylsulfanyl-butyric acid methyl ester-2yl-carbamoyl;(2S),4-methylsulfanyl-butyric acid-2yl-carbamoyl; phenoxy.

For p: 1-2, especially 2; a further preferred value is 0.

For L: —C(O)—NH—; —CH₂—C(O)—NH—; —CH₂—NH—C(O)—; —CH₂—NH—SO₂—; especially—C(O)—NH—.

For A: phenyl; pyridyl, thienyl; naphthyl.

For R¹⁶ & R¹⁸⁻²⁶: H, C₁₋₄alkyl, especially H.

In another embodiment of the invention preferred values are set outbelow.

In compounds of Formula III: X¹ is H or methoxyC₁₋₄alkyl (especially H);X² is H. phenyl or benzyl (especially benzyl); X³ is H or C₁₋₄alkyl(especially H); X⁴ is C₁₋₄alkylsulfanyl (especially methylsulfanyl); andA is phenyl. When A is a 6-membered aryl or heteroaryl ring then groups—NX¹— and the substituent comprising X⁴ are preferably in metajuxtaopsition relative to each other; and X², if present, is preferablypositioned para relative to —NX¹—. The chiral carbon to which —COOX³ isattached is preferably in S configuration. The chiral carbons at the 2and 4 positions of the pyrrolidine ring are preferably in Sconfiguration.

In compounds of Formula IV: X⁵ is —CO—C₁₋₄alkyl (especially—CO—CH₂—CHMe₂) or —CH₂—Ph—O—C₁₋₄alkyl (especially —CH₂—Ph—OMe);heteroaryl is preferably pyridyl and a preferred aryl or heteroarylsubstituent is —O—C₁₋₄alkyl (especially methoxy); and A is naphthyl. Thechiral carbons at the 2 and 4 positions of the pyrrolidine ring arepreferably in S configuration. The attachment point for A relative to—(CH₂)_(1,2)— is preferably at the 1 position of napththalene and theequivalent position for heterocyclic values for A (regardless of ringnumbering conventions for heterocycles). A preferred value for—(CH₂)_(1,2)— is —(CH₂)₂—.

In compounds of Formula V: X⁶ is —CO—C₁₋₅alkyl (more preferably—CO—CH₂—CHMe₂ or —CO—CH₂-t-butyl, especially —CO—CH₂—CHMe₂) or—CH₂—Ph—O—C₁₋₄alkyl (especially —CH₂—Ph—OMe); heteroaryl is preferablypyridyl and a preferred aryl substitution is —O—C₁₋₄alkyl (especiallymethoxy); and A is phenyl or naphthyl (especially phenyl). The chiralcarbons at the 2 and 4 positions of the pyrrolidine ring are preferablyin S configuration. A preferred value for —(CH₂)_(1,2)— is —(CH₂)₁—.

Suitable pairs of values for R³ when p=2 are: —COOMe, —CO,N(Me).OMe;NO₂, —CO.N(Me).OMe; —COOMe, allyloxycarbonyl; —CO.N(Me).OMe,allyloxycarbonyl; allyloxycarbonyl, —CO.N(Me).O.CH₂CH═CH₂; OH, COOH;—COOMe, COOMe; Ph, —CO.N-Methionine methyl ester; Ph, —CO.N-Methionine;benzyl, —CO.N-Methionine methyl ester; benzyl, —CO.N-Methionine; benzyl,—CO.N-Methionine isopropyl ester; Ph, —CO.Nα-Glutamine methyl ester; Ph,—CO.Nα-Glutamine.

Suitable values for L=CHNR²¹T include CH₂.N(CO.CH₂.CHMe₂).CH₂.CH₂;CH₂.N(CH₂CH₂CH₂OMe).CH₂.CH₂; CH₂.N(CH₂.pPh.OMe).CH₂.CH₂;CH₂.N(CO.CH₂.CHMe₂).CH₂; CH₂N(CO.CH₂.CH₂.CH₂.Me).CH₂;CH₂N(CO.CH₂.CHMe.CH₂Me).CH₂; CH₂N(CO.CH₂.CH₂.OMe)CH₂;CH₂N(CO.CH₂.pyridin-3-yl).CH₂; CH₂N(4-methoxybenzyl)CH₂;CH₂N(CO.CH₂.CHMe₂)CH₂.CH₂.CH(Ph); CH₂N(CO.CH₃)CH₂.CH₂.CH(Ph);CH₂N(CO.CH₂.CHMe₂)CH₂; CH₂N(CO.CH₃)CH₂; CH₂N(CO.CH₂.CHMe₂)CH₂.CH(Ph);CH₂N(CO.CH₂.CMe₃)CH₂.CH(Ph); CH₂N(CO.CH₂.pyridin-3-yl)CH₂.CH(Ph);CH₂N(CO.1-hydroxy-6methoxy-pyridin-3-yl)CH₂.CH(Ph);CH₂N(CO.CH₂CHMe)CH₂.CH₂; CH₂N(CO.CH₂CMe₃)CH₂.CH₂;CH₂N(CO.CH₂pyridin-3-yl)CH₂.CH₂; CH₂N(CO.4-methoxybenzyl)CH₂.CH₂;

Suitable values for L=—CH₂NR¹⁸— include CH₂NH; CH₂NMe;CH₂N(CO.CH₂.CHMe₂) and CH₂N(CO.CH₂.CH₂.OMe).

Various forms of prodrugs are well known in the art. For examples ofsuch prodrug derivatives, see:

a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) andMethods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al.(Academic Press. 1985);

b) A Textbook of Drug Design and Development, edited byKrogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and Application ofProdrugs”, by H. Bundgaard p. 113-191 (1991);

c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);

d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285(1988); and

e) N. Kakeya, et al., Chem Pharm Bull, 32, 692 (1984).

Examples of prodrugs include in vivo hydrolysable esters of a compoundof the Formula I. An in vivo hydrolysable ester of a compound of theformula (I) containing carboxy group is, for example, apharmaceutically-acceptable ester which is hydrolysed in the human oranimal body to produce the parent acid. Suitablepharmaceutically-acceptable esters for carboxy include C₁₋₆alkoxymethylesters for example methoxymethyl, C₁₋₆alkanoyloxymethyl esters forexample pivaloyloxymethyl, phthalidyl esters.C₃₋₈cycloalkoxycarbonyloxyC₁₋₆alkyl esters for example1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters forexample 5-methyl-1,3-dioxolen-2-onylmethyl; andC₁₋₆alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethyland may be formed at any carboxy group in the compounds of thisinvention.

Particular substitutions on A for 6 membered rings are in the meta orpara positions.

Some compounds within the scope of Formula I are known as intermediatesin carbapenem side chain synthesis but it is believed that they have notbeen previously described in forms suitable as pharmaceuticalcompositions nor had any pharmaceutical activity associated with themper se. The reader is referred to the following publications in thisregard and also in respect of synthetic details for compoundpreparation: Matsumura, Heterocycles (1995), 41, 147-59; European patentapplication EP 590885 (Zeneca; Betts et al); European patent applicationEP 592167 (Zeneca; Siret); European patent application EP 562855(Zeneca; Jung et al); International patent application WO 92/17480(Imperial Chemical Industries; Betts et al); European patent applicationEP 508682 (Imperial Chemical Industries; Betts et al); European PatentApplication EP 280771 (Fujisawa Pharmaceutical, Murata et al): andInternational patent application WO 92/17479 (Imperial ChemicalIndustries; Betts et al).

In this specification the generic term “alkyl” includes bothstraight-chain and branched-chain alkyl groups. However references toindividual alkyl groups such as “propyl” are specific for thestraight-chain version only and references to individual branched-chainalkyl groups such as “isopropyl” are specific for the branched-chainversion only. An analogous convention applies to other generic terms.

It is to be understood that, insofar as certain of the compounds ofFormula I defined above may exist in optically active or racemic formsby virtue of one or more asymmetric carbon atoms, the invention includesin its definition any such optically active or racemic form whichpossesses the property of inhibiting FTPase. The synthesis of opticallyactive forms may be carried out by standard techniques of organicchemistry well known in the art, for example by synthesis from opticallyactive starting materials or by resolution of a racemic form. Similarly,inhibitory properties against FTPase may be evaluated using the standardlaboratory techniques referred to hereinafter.

The term “halogen” refers to fluorine, chlorine, bromine and iodine. Theterm “carbamoyl” refers to —C(O)NH₂. The term “BOC” refers totert-butyl-O—C(O)—. The term “allyl” refers to CH₂═CH—CH₂—. Bicyclicaryl and bicyclic heteroaryl rings refer to ring systems in which bothrings of the bicyclic system are aromatic.

Examples of C₁₋₆alkyl include methyl, ethyl, propyl, isopropyl,sec-butyl, tert-butyl and pentyl; examples of C₁₋₄alkyl include methyl,ethyl, propyl, isopropyl, sec-butyl and tert-butyl; examples ofC₁₋₃alkyl include methyl, ethyl, propyl and isopropyl; examples of—C₁₋₃alkylenePh include benzyl, phenylethyl, phenylpropyl; examples ofC₁₋₄alkoxy (also called —O—C₁₋₄alkyl herein) include methoxy, ethoxy andpropoxy; examples of C₁₋₄alkanoyl include formyl, acetyl and propionyl;examples of C₁₋₄alkanoyloxy include acetyloxy and propionyloxy; examplesof C₁₋₄alkylamino include methylamino, ethylamino, propylamino,isopropylamino, sec-butylamino and tert-butylamino; examples ofdi-(C₁₋₄alkyl)amino include di-methylamino, di-ethylamino andN-ethyl-N-methylamino; examples of C₁₋₄alkanoylamino include acetamidoand propionylamino; examples of C₁₋₄alkoxycarbonyl includemethoxycarbonyl, ethoxycarbonyl and propoxycarbonyl; examples ofC₁₋₄alkylsulfanyl include methylsulfanyl, ethylsulfanyl, propylsulfanyl,isopropylsulfanyl sec-butylsulfanyl and tert-butylsulfanyl; examples ofC₁₋₄alkylsulfinyl include methylsulfinyl, ethylsulfinyl, propylsulfinyl,isopropylsulfinyl, sec-butylsulfinyl and tert-butylsulfinyl; examples ofC₁₋₄alkylsulfonyl include methylsulfonyl, ethylsulfonyl, propylsulfonyl,isopropylsulfonyl, sec-butylsulfonyl and tert-butylsulfonyl; examples of—CO—C₁₋₄alkyl include formyl, acetyl, propionyl, butyryl, and valeryl;examples of —CO—O—C₁₋₄alkyl include ethyloxycarbonyl; propyloxycarbonyland tert-butyloxycarbonyl (BOC); examples of —CO—O—C₂₋₄alkenyl includeallyloxycarbonyl and vinyloxycarbonyl; examples of —CO—O—(CH₂)_(n)Phwhere n=0-4 include phenyloxycarbonyl, benzyloxycarbonyl,phenylethyloxycarbonyl and phenylpropyloxycarbonyl; examples of—C₁₋₄alkylene-CONR⁴R⁵ include carbamoylmethyl, carbamoylethyl,N-methylcarbamoylethyl, N-methyl-N-ethylcarbamoylethyl; examples of—C₁₋₄alkylene-COOR⁶ include carboxymethyl, carboxyethyl, carboxypropyl,propionic acid methyl ester, acetic acid ethyl ester; examples ofC₂₋₄alkenyl include allyl and vinyl; examples of —O—C₂₋₄alkenyl includeallyloxy and vinyloxy; examples of lipophilic amino acids includevaline, leucine, isoleucine, methionine, phenylalanine, serine,threonine and tyrosine; examples of carbamoylC₁₋₄alkyl includecarbamoylmethyl, carbamoylethyl and carbamoylpropyl; examples ofN-(monoC₁₋₄alkyl)carbamoylC₁₋₄alkyl include N-methyl-carbamoylmethyl andN-ethyl-carbamoylethyl; examples of N-(diC₁₋₄alkyl)carbamoyl-C₁₋₄alkylinclude N,N-dimethylcarbamoylethyl and N-methyl-N-ethylcarbamoylethyl;examples of C₁₋₄alkyl monosubstituted on carbon with ═N—OH includebutyraldehyde oxime and propionaldehyde oxime; examples ofhydroxyC₁₋₆alkyl include hydroxymethyl, hydroxyethyl, hydroxypropyl,2-hydroxypropyl, 2-(hydroxymethyl)propyl and hydroxypentyl; examples ofC₁₋₆alkoxyC₁₋₆alkyl include methoxyethyl, ethoxyethyl and methoxybutyl;examples of C₁₋₆alkylcarbonyl include methylcarbonyl, ethylcarbonyl,propylcarbonyl, isopropylcarbonyl, sec-butylcarbonyl, tert-butylcarbonyland pentylcarbonyl; examples of hydroxyC₁₋₆alkylcarbonyl includehydroxyacetyl, hydroxypropionyl, hydroxybutyryl, 3-hydroxybutyryl andhydroxypentanoyl; examples of C₁₋₆alkoxyC₁₋₆alkylcarbonyl includemethoxyacetyl, methoxypropionyl, ethoxybutyryl and butoxyacetyl;examples of phenylC₁₋₆alkyl include benzyl, phenylethyl andphenylpropyl; examples of —CO—C₁₋₄alkyl-Ph include phenylacetyl andphenylpropionyl; examples of —CO—C₁₋₄alkyl-heteroaryl include2-(3-pyridyl)-acetyl and 2-(3-thienyl)-acetyl; examples ofN-(C₁₋₆alkyl)carbamoyl include N-methyl-carbamoyl and N-ethyl-carbamoyl;examples of N-(diC₁₋₆alkyl)carbamoyl include-N,N-dimethylcarbamoyl andN-methyl-N-ethylcarbamoyl.

Examples of 5-10 membered monocyclic or bicyclic heteroaryl ringscontaining upto 5 heteroatoms selected from O,N and S include thefollowing. Examples of 5- or 6-membered heteroaryl ring systems includeimidazole, triazole, pyrazine, pyrimidine, pyridazine, pyridine,isoxazole, oxazole, isothiazole, thiazole and thiophene. A 9 or 10membered bicyclic heteroaryl ring system is an aromatic bicyclic ringsystem comprising a 6-membered ring fused to either a 5 membered ring oranother 6 membered ring. Examples of 5/6 and 6/6 bicyclic ring systemsinclude benzofuran, benzimidazole, benzthiophene, benzthiazole,benzisothiazole, benzoxazole, benzisoxazole, pyridoimidazole,pyrimidoimidazole, quinoline, isoquinoline, quinoxaline, quinazoline,phthalazine, cinnoline and naphthyridine.

Preferably monocyclic heteroaryl rings contain upto 3 heteroatoms andbicyclic heteroaryl rings contain upto 5 heteroatoms. Preferredheteroatoms are N and S, especially N. In general, attachment ofheterocyclic rings to other groups is via carbon atoms. Suitable valuesof heterocycles containing only N as the heteroatom are pyrrole,pyridine, indole, quinoline, isoquinoline, imidazole, pyrazine,pyrimidine, purine and pteridine.

Preferably any chiral carbon atoms at the 2 and 4 positions of thepyrrolidine ring in Formulas I and III-V are in S configuration.

Compounds of Formula I and III-V may form salts which are within theambit of the invention. Pharmaceutically acceptable salts are preferredalthough other salts may be useful in, for example, isolating orpurifying compounds.

When the compound contains a basic moiety it may form pharmaceuticallyacceptable salts with a variety of inorganic or organic acids, forexample hydrochloric, hydrobromic, sulphuric, phosphoric,trifluoroacetic, citric or maleic acid. A suitablepharmaceutically-acceptable salt of the invention when the compoundcontains an acidic moiety is an alkali metal salt, for example a sodiumor potassium salt, an alkaline earth metal salt, for example a calciumor magnesium salt, an ammonium salt or a salt with an organic base whichaffords a pharmaceutically-acceptable cation, for example a salt withmethylamine, dimethylamine, trimethylamine, piperidine, morpholine ortris-(2-hydroxyethyl)amine.

Solvates, for example hydrates, are also within the ambit of theinvention and may be prepared by generally known methods.

According to another aspect of the present invention there is provided acompound of Formula I for use as a medicament.

According to another aspect of the present invention there is providedthe use of a compound of Formula I in preparation of a medicament fortreating ras mediated diseases, especially cancer.

According to another aspect of the present invention there is provided amethod of treating ras mediated diseases, especially cancer, byadministering an effective amount of a compound of Formula I to a mammalin need of such treatment.

According to a further feature of the invention there is provided acompound of Formula I, or a pharmaceutically-acceptable salt thereof,for use in a method of treatment of the human or animal body by therapy.

The invention also includes a method of treating a disease or medicalcondition mediated alone or in part by farnesylated ras which comprisesadministering to a mammal requiring such treatment an effective amountof an active ingredient as defined above. The invention also providesthe use of such an active ingredient in the production of a newmedicament for use in a farnesylated ras mediated disease or medicalcondition.

Specific cancers of interest include:

carcinoma, including that of the bladder, breast, colon, kidney, liver,lung, ovary, pancreas, stomach, cervix, thyroid and skin;

hematopoietic tumors of lymphoid lineage, including acute lymphocyticleukemia, B-cell lymphoma and Burketts lymphoma;

hematopoietic tumors of myeloid lineage, including acute and chronicmyelogenous leukemias and promyelocytic leukemia;

tumors of mesenchymal origin, including fibrosarcoma andrhabdomyosarcoma; and

other tumors, including melanoma, seminoma, tetratocarcinoma,neuroblastoma and glioma.

The compounds of Formula I are especially useful in treatment of tumorshaving a high incidence of ras mutation, such as colon, lung, andpancreatic tumors. By the administration of a composition having one (ora combination) of the compounds of this invention, development of tumorsin a mammalian host is reduced.

Compounds of Formula I may also be useful in the treatment of diseasesother than cancer that may be associated with signal transductionpathways operating through Ras. e.g., neuro-fibromatosis.

Compounds of Formula I may also be useful in the treatment of diseasesassociated with CAAX-containing proteins other than Ras (e.g., nuclearlamins and transducin) that are also post-translationally modified bythe enzyme farnesyl protein transferase.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions. dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insufflation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, intramuscular orintramuscular dosing or as a suppository for rectal dosing).

The compositions of the invention may be obtained by conventionalprocedures using conventional pharmaceutical excipients, well known inthe art. Thus, compositions intended for oral use may contain, forexample, one or more colouring, sweetening, flavouring and/orpreservative agents.

Suitable pharmaceutically acceptable excipients for a tablet formulationinclude, for example, inert diluents such as lactose, sodium carbonate,calcium phosphate or calcium carbonate, granulating and disintegratingagents such as corn starch or algenic acid; binding agents such asstarch; lubricating agents such as magnesium stearate, stearic acid ortalc; preservative agents such as ethyl or propyl p-hydroxybenzoate, andanti-oxidants, such as ascorbic acid. Tablet formulations may beuncoated or coated either to modify their disintegration and thesubsequent absorption of the active ingredient within thegastrointestinal tract, or to improve their stability and/or appearance,in either case, using conventional coating agents and procedures wellknown in the art.

Compositions for oral use may be in the form of hard gelatin capsules inwhich the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the active ingredient is mixed with water oran oil such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions generally contain the active ingredient in finelypowdered form together with one or more suspending agents, such assodium carboxymethylcellulose methylcellulose,hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone,gum tragacanth and gum acacia; dispersing or wetting agents such aslecithin or condensation products of an alkylene oxide with fatty acids(for example polyoxethylene stearate), or condensation products ofethylene oxide with long chain aliphatic alcohols, for exampleheptadecaethyleneoxycetanol, or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with long chain aliphatic alcohols, for exampleheptadecaethyleneoxycetanol, or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives (such as ethyl orpropyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid),colouring agents, flavouring agents, and/or sweetening agents (such assucrose, saccharine or aspartame).

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil (such as arachis oil, olive oil, sesame oil orcoconut oil) or in a mineral oil (such as liquid paraffin). The oilysuspensions may also contain a thickening agent such as beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set outabove, and flavouring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspending agentand one or more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients such as sweetening, flavouring and colouringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, or a mineral oil, such as for exampleliquid paraffin or a mixture of any of these. Suitable emulsifyingagents may be, for example, naturally-occurring gums such as gum acaciaor gum tragacanth naturally-occurring phosphatides such as soya bean,lecithin, an esters or partial esters derived from fatty acids andhexitol anhydrides (for example sorbitan monooleate) and condensationproducts of the said partial esters with ethylene oxide such aspolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening, flavouring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents such asglycerol, propylene glycol, sorbitol, aspartame or sucrose, and may alsocontain a demulcent, preservative, flavouring and/or colouring agent.

The pharmaceutical compositions may also be in the form of a sterileinjectable aqueous or oily suspension, which may be formulated accordingto known procedures using one or more of the appropriate dispersing orwetting agents and suspending agents, which have been mentioned above. Asterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally-acceptable diluent or solvent,for example a solution in 1,3-butanediol.

Suppository formulations may be prepared by mixing the active ingredientwith a suitable non-irritating excipient which is solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum to release the drug. Suitable excipients include, forexample, cocoa butter and polyethylene glycols.

Topical formulations, such as creams, ointments, gels and aqueous oroily solutions or suspensions, may generally be obtained by formulatingan active ingredient with a conventional, topically acceptable, vehicleor diluent using conventional procedure well known in the art.

Compositions for administration by insufflation may be in the form of afinely divided powder containing particles of average diameter of, forexample, 30μ or much less. the powder itself comprising either activeingredient alone or diluted with one or more physiologically acceptablecarriers such as lactose. The powder for insufflation is thenconveniently retained in a capsule containing, for example, 1 to 50 mgof active ingredient for use with a turbo-inhaler device, such as isused for insufflation of the known agent sodium cromoglycate.

Compositions for administration by inhalation may be in the form of aconventional pressurised aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient.

For further information on Formulation the reader is referred to Chapter25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch;Chairman of Editorial Board). Pergamon Press 1990.

The amount of active ingredient that is combined with one or moreexcipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. For example, a formulation intended for oraladministration to humans will generally contain, for example, from 0.5mg to 2 g of active agent compounded with an appropriate and convenientamount of excipients which may vary from about 5 to about 98 percent byweight of the total composition. Dosage unit forms will generallycontain about 1 mg to about 500 mg of an active ingredient. For furtherinformation on Routes of Administration and Dosage Regimes the reader isreferred to Chapter 25.3 in Volume 5 of Comprehensive MedicinalChemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press1990.

The size of the dose for therapeutic or prophylactic purposes of acompound of the Formula I will naturally vary according to the natureand severity of the conditions, the age and sex of the animal or patientand the route of administration, according to well known principles ofmedicine. As mentioned above, compounds of the Formula I are useful intreating diseases or medical conditions which are due alone or in partto the effects of farnesylation of ras.

In using a compound of the Formula I for therapeutic or prophylacticpurposes it will generally be administered so that a daily dose in therange, for example, 0.5 mg to 75 mg per kg body weight is received,given if required in divided doses. In general lower doses will beadministered when a parenteral route is employed. Thus, for example, forintravenous administration, a dose in the range, for example, 0.5 mg to30 mg per kg body weight will generally be used. Similarly, foradministration by inhalation, a dose in the range, for example, 0.5 mgto 25 mg per kg body weight will be used. Oral administration is howeverpreferred.

Compounds of this invention may be useful in combination with knownanti-cancer and cytotoxic agents. If formulated as a fixed dose suchcombination products employ the compounds of this invention within thedosage range described herein and the other pharmaceutically activeagent within its approved dosage range. Sequential use is contemplatedwhen a combination formulation is inappropriate.

Although the compounds of the Formula I are primarily of value astherapeutic agents for use in warm-blooded animals (including man), theyare also useful whenever it is required to inhibit the effects ofactivation of ras by farnesylation. Thus, they are useful aspharmacological standards for use in the development of new biologicaltests and in the search for new pharmacological agents.

According to another aspect of the present invention there is providedindividual compounds produced as end products in the Examples set outbelow and salts thereof.

A compound of the invention, or a salt thereof, may be prepared by anyprocess known to be applicable to the preparation of such compounds orstructurally related compounds. Such processes are illustrated by thefollowing representative schemes in which variable groups have any ofthe meanings defined for Formula I unless stated otherwise. Functionalgroups may be protected and deprotected using conventional methods. Forexamples of protecting groups such as amino and carboxylic acidprotecting groups (as well as means of formation and eventualdeprotection), see T. W. Greene and P. G. M. Wuts, “Protective Groups inOrganic Synthesis”, Second Edition, John Wiley & Sons, New York, 1991.Note abbreviations used have been listed immediately before the Examplesbelow.

Protecting groups may be removed by any convenient method as describedin the literature or known to the skilled chemist as appropriate for theremoval of the protecting group in question, such methods being chosenso as to effect removal of the protecting group with minimum disturbanceof groups elsewhere in the molecule.

Specific examples of protecting groups are given below for the sake ofconvenience, in which “lower” signifies that the group to which it isapplied preferably has 1-4 carbon atoms. It will be understood thatthese examples are not exhaustive. Where specific examples of methodsfor the removal of protecting groups are given below these are similarlynot exhaustive. The use of protecting groups and methods of deprotectionnot specifically mentioned is of course within the scope of theinvention.

A carboxy protecting group may be the residue of an ester-formingaliphatic or araliphatic alcohol or of an ester-forming silanol (thesaid alcohol or silanol preferably containing 1-20 carbon atoms).

Examples of carboxy protecting groups include straight or branched chain(1-12C)alkyl groups (e.g. isopropyl, t-butyl); lower alkoxy lower alkylgroups (e.g. methoxymethyl, ethoxymethyl, isobutoxymethyl; loweraliphatic acyloxy lower alkyl groups, (e.g. acetoxymethyl,propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl); loweralkoxycarbonyloxy lower alkyl groups (e.g. 1-methoxycarbonyloxyethyl,1-ethoxycarbonyloxyethyl); aryl lower alkyl groups (e.g.p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, benzhydryl andphthalidyl); tri(lower alkyl)silyl groups (e.g. trimethylsilyl andt-butyldimethylsilyl); tri(lower alkyl)silyl lower alkyl groups (e.g.trimethylsilylethyl); and (2-6C)alkenyl groups (e.g. allyl andvinylethyl).

Methods particularly appropriate for the removal of carboxyl protectinggroups include for example acid-, metal- or enzymically-catalysedhydrolysis.

Examples of hydroxy protecting groups include lower alkenyl groups (e.g.allyl); lower alkanoyl groups (e.g. acetyl); lower alkoxycarbonyl groups(e.g. t-butoxycarbonyl); lower alkenyloxycarbonyl groups (e.g.allyloxycarbonyl); aryl lower alkoxycarbonyl groups (e.g.benzoyloxycarbonyl, p-methoxybenzyloxycarbonyl,o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl); tri loweralkyl/arylsilyl groups (e.g. trimethylsilyl, t-butyldimethylsilyl,t-butyldiphenylsilyl); aryl lower alkyl groups (e.g. benzyl) groups; andtriaryl lower alkyl groups (e.g. triphenylmethyl).

Examples of amino protecting groups include formyl, aralkyl groups (e.g.benzyl and substituted benzyl, e.g. p-methoxybenzyl, nitrobenzyl and2,4-dimethoxybenzyl, and triphenylmethyl); di-p-anisylmethyl andfurylmethyl groups; lower alkoxycarbonyl (e.g. t-butoxycarbonyl); loweralkenyloxycarbonyl (e.g. allyloxycarbonyl); aryl lower alkoxycarbonylgroups (e.g. benzyloxycarbonyl, p-methoxybenzyloxycarbonyl,o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl; trialkylsilyl (e.g.trimethylsilyl and t-butyldimethylsilyl): alkylidene (e.g. methylidene);benzylidene and substituted benzylidene groups.

Methods appropriate for removal of hydroxy and amino protecting groupsinclude, for example, acid-, base, metal- or enzymically-catalysedhydrolysis, or photolytically for groups such aso-nitrobenzyloxycarbonyl, or with fluoride ions for silyl groups.

Examples of protecting groups for amide groups include aralkoxymethyl(e.g. benzyloxymethyl and substituted benzyloxymethyl); alkoxymethyl(e.g. methoxymethyl and trimethylsilylethoxymethyl); tri alkyl/arylsilyl(e.g. trimethylsilyl, t-butyldimethylsily, t-butyldiphenylsilyl); trialkyl/arylsilyloxymethyl (e.g. t-butyldimethylsilyloxymethyl,t-butyldiphenylsilyloxymethyl); 4-alkoxyphenyl (e.g. 4-methoxyphenyl);2,4-di(alkoxy)phenyl (e.g. 2,4dimethoxyphenyl); 4-alkoxybenzyl (e.g.4-methoxybenzyl); 2,4-di(alkoxy)benzyl (e.g. 2,4-di(methoxy)benzyl); andalk-1-enyl (e.g. allyl, but-1-enyl and substituted vinyl e.g.2-phenylvinyl).

Aralkoxymethyl, groups may be introduced onto the amide group byreacting the latter group with the appropriate aralkoxymethyl chloride,and removed by catalytic hydrogenation, Alkoxymethyl, trialkyl/arylsilyl and tri alkyl/silyloxymethyl groups may be introduced byreacting the amide with the appropriate chloride and removing with acid;or in the case of the silyl containing groups, fluoride ions. Thealkoxyphenyl and alkoxybenzyl groups are conveniently introduced byarylation or alkylation with an appropriate halide and removed byoxidation with ceric ammonium nitrate. Finally alk-1-enyl groups may beintroduced by reacting the amide with the appropriate aldehyde andremoved with acid.

Compounds of Formula I in which L represents —CO—NR¹⁶— may be preparedby forming an amide bond between compounds 1 and 2 as outlined in Scheme23. Compounds of Formula I in which L represents —CO—NR²⁵—T— may beprepared by an analogous procedure. Suitable coupling conditions includethe following.

i) Use of EEDQ at room temperature in an organic solvent (e.g.dichloromethane, methanol).

ii) Use of oxalyl chloride in an organic solvent (e.g. DMF, CH₂Cl₂) inthe presence of an organic base (e.g. NMM, triethylamine, DMAP) at 0° toroom temperature for 0.5-16 h.

iii) Use of EDC/HOBT in an organic solvent (e.g. DMF, CH₂Cl₂).

iv) Use of DCCI/HOBT in an organic solvent (e.g. DMF, CH₂Cl₂) in thepresence of an organic base (e.g. triethylamine).

v) Use of mixed anhydride reactions under standard conditions, forexample isopropylchloroformate in an organic solvent (e.g. DMF, DMA,dichloromethane) in the presence of an organic base (e.g. NMM, DMAP,triethylamine).

vi) Via an active ester under standard conditions e.g. pentafluorophenylester in an organic solvent (e.g. dichloromethane) in the presence of anorganic base (e.g. triethylamine).

vii) Via an acid chloride under standard conditions e.g. using thionylchloride and heat for about 150 min followed by an organic base (e.g.triethylamine) in the presence of an organic solvent (e.g.acetonitrile).

Compounds of Formula I in which L represents —CH₂NR¹⁸—, —CH₂O— or —CH₂S—may be prepared as outlined in Scheme 24. LG represents a leaving group(e.g. mesyloxy, tosyloxy, halogen) and X represents S. O or NR¹⁸.Suitable coupling conditions include the following.

i) Use of an inorganic base (e.g. NaHCO₃, NaH, K₂CO₃, butyllithium) inan organic solvent (e.g. THF, DMF, DMSO) and a temperature of about 70°to 150°

ii) Use of an organic base (e.g. triethylamine, DMAP) in an organicsolvent (e.g. THF, dichloromethane, DMA, DMF) at a temperature range ofroom temperature −150°

iii) Use of an inorganic base (e.g. KOH, NaOH, K₂CO₃) in an aqueous(e.g. water) and organic solvents (e.g. dichloromethane) in a 2 phasesystem, optionally in the presence of a phase transfer catalyst (e.g.tetrabutylammoniumbromide).

Compounds of Formula I in which L represents —CH═CR²⁰— may be preparedusing a Wittig reaction as outlined in Scheme 25. Suitable reactionconditions include the following.

i) Use of a base (e.g. potassium carbonate, metal hydride, metalalkoxide) in the presence of an organic solvent (e.g. THF, toluene,DMSO) optionally in the presence of an aqueous solvent (2-phase system)and optionally in the presence of a catalyst complexing agent whichsolubilises alkali metal ions in non-polar solvents such as1,4,7,10,13-pentaoxacyclopentadecane (also called 15-Crown-5) or1,4,7,10,13,16-hexaoxacyclooctadecane (also called 18-Crown-6).

Compounds of Formula I in which L represents —CH₂—NR¹⁸— may be preparedas outlined in Scheme 26 by coupling aldehyde (2) with compound 4.Suitable coupling conditions include the following.

i) Use of a reducing agent (e.g. NaCNBH₃, BH₃, hydrogen plus catalyst,LiHBEt₃, di-isobutyl-aluminiumhydride, lithium aluminium hydride, sodiumborohydride) in the presence of a suitable solvent e.g. ethanol & aceticacid.

Aldehyde (2) may be prepared by oxidation of the corresponding alcohol(1) under suitable conditions such as use of an oxidising agent (e.g.TPAP, NMM-O) in the presence of an organic solvent (e.g. acetonitrile,dichloromethane) at room temperature. Other suitable oxidising agentsinclude chromium oxide, pyridinium chlorochromate, pyridiniumdichromate, sodium dichromate and sodium hypochlorite.

Aldehyde (2) may also be prepared by reduction of the correspondingester (1) under standard conditions using for examplediisobutyl-aluminium hydride.

Compounds of Formula I in which L represents —CH₂—NR²¹—T—, —CH₂—O—T— or—CH₂—S—T— may be prepared as outlined in Scheme 27 in which LGrepresents a leaving group (e.g. mesyloxy, tosyloxy, halogen) and Xrepresents O, S or NR²¹. Suitable coupling conditions are as outlinedabove in relation to Scheme 24. Optionally the positions of LG and XH incompounds 1 & 2 in Scheme 27 can be reversed to give the same endproduct.

Compounds of Formula I in which L represents —CH₂—NR²³—SO_(2—) may beprepared as outlined in Scheme 28. Compounds 1 & 2 may be coupled understandard conditions such as the following.

i) Use of an organic base (e.g. di-isopropyl-ethylamine, triethylamine,4-methyl-morpholine) in the presence of an organic solvent (e.g.dichloromethane) at a temperature range of 0°-40°

ii) Use of an inorganic base (e.g. potassium carbonate) in the presenceof an organic solvent (e.g. DMF) at a temperature range of 0°-150°

Compounds of Formula I in which L represents —CH₂—NR²⁴—CO—T— may beprepared as outlined in Scheme 29. Compounds 1 & 2 may be coupled understandard conditions such as described above for L=—CO—NR¹⁶—.

Compounds of Formula I in which L represents —CH₂—CHR¹⁹— may be preparedas by reduction of compounds of the type set out as compound 3 in Scheme25 but substituting R¹⁹ in lieu of R²⁰. Reduction is carried out understandard conditions with standard reagents for example usinghydrogenation in the presence of a catalyst such as palladium oncharcoal at room temperature.

Biological activity was tested as follows. Farnesyl protein transferase(FPT) was partially purified from human placenta by ammonium sulphatefractionation followed by a single Q-Sepharose® (Pharmacia, Inc) anionexchange chromatography essentially as described by Ray andLopez-Belmonte (Ray K P and Lopez-Belmonte J (1992) Biochemical SocietyTransations 20 494-497). The substrate for FPT was Kras (CVIM C-terminalsequence). The cDNA for oncogenic val12 variant of human c-Ki-ras-2 4Bwas obtained from the plasmid pSW11-1 (ATCC). This was then subclonedinto the polylinker of a suitable expression vector e.g. pIC147. TheKras was obtained after expression in the E. coli strain, BL21. Theexpression and purification of c-KI-ras-2 4B and the val12 variant in E.coli has also been reported by Lowe et al (Lowe P N et al. J. Biol.Chem. (1991) 266 1672-1678).

Incubations with enzyme contained 300 nM tritiated farnesylpyrophosphate (DuPont/New England Nuclear), 120 nM ras-CVIM, 50 mM TrisHCl pH 8.0, 5 mM MgCl₂, 10 μM ZnCl₂, 5 mM dithiotheitol and compoundswere added at appropriate concentrations in DMSO (3% final concentrationin test and vehicle control). Incubations were for 20 minutes at 37° andwere stopped with acid ethanol as described by Pompliano et al.(Pompliano D L et al (1992) 31 3800-3807). Precipitated protein was thencollected onto glass fibre filter mats (B) using a Tomtec® cellharvester and tritiated label was measured in a Wallac®1204 Betaplatescintillation counter.

Although the pharmacological properties of the compounds of the FormulaI vary with structural change as expected, in general compounds of theFormula I possess an IC₅₀ in the above test in the range, for example,0.01 to 200 μM. Thus by way of example, the compound5{[(2S,4S),4-acetylsulfanyl-1-(4-nitro-benzyloxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-3(N-methyl-methoxycarbamoyl)-benzoic acid allyl ester (see Example 7)has an IC₅₀ of approximately 0.5 μM. No physiologically unacceptabletoxicity was observed at the effective dose for compounds tested of thepresent invention.

The invention will now be illustrated in the following non-limitingExamples in which, unless otherwise stated:

(i) evaporations were carried out by rotary evaporation in vacuo andwork-up procedures were carried out after removal of residual solids byfiltration;

(ii) operations were carried out at room temperature, that is in therange 18-25° C. and under an atmosphere of an inert gas such as argon;

(iii) column chromatography (by the flash procedure) and medium pressureliquid chromatography (MPLC) were performed on Merck Kieselgel silica(Art. 9385) or Merck Lichroprep RP-18 (Art. 9303) reversed-phase silicaobtained from E. Merck, Darmstadt, Germany;

(iv) yields are given for illustration only and are not necessarily themaximum attainable;

(v) the end-products of the Formula I have satisfactory microanalysesand their structures were confirmed by nuclear magnetic resonance (NMR)and mass spectral techniques; chemical shift values were measured on thedelta scale; the following abbreviations have been used; s, singlet; d,doublet; t or tr, triplet; m, multiplet; br, broad;

(vi) intermediates were not generally fully characterised and purity wasassessed by thin layer chromatographic, infra-red (IR) or NMR analysis;

(vii) melting points are uncorrected and were determined using a MettlerSP62 automatic melting point apparatus or an oil-bath apparatus; meltingpoints for the end-products of the Formula I were determined aftercrystallisation from a conventional organic solvent such as ethanol,methanol, acetone, ether or hexane, alone or in admixture; and

(viii) the following abbreviations have been used:

BOC tert-butoxycarbonyl

DCCI 1,3-dicyclohexylcarbodiimide

DMA N,N-dimethylacetamide

DMAP 4-dimethyl-aminopyridine

DMF N,N-dimethylformamide

DMSO dimethylsulfoxide

EDC 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide

EEDQ 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline

HOBT 1-hydroxybenzotriazole

NMM N-methylmorpholine

NMM-O 4-methylmorpholine-N-oxide

TFA trifluoroacetic acid

THF tetrahydrofuran

TMSI trimethylsilyliodide

TPAP tetrapropylammonium perruthenate

Note in the Schemes only those hydrogen atoms thought to assist clarityhave been illustrated (ie not all hydrogen atoms have been illustrated).

EXAMPLE 1

(See Scheme 1)

(2S,4S)-4-acetylsulfanyl-2[3nitro-5-(N-methoxy-N-methyl-carbamoyl)-phenylcarbamoyl]-pyrrolidine-1-carboxylicacid 4-nitro-benzyl ester

A mixture of 4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid1-(4-nitrobenzyl) ester (1(c)) (0.2 g) and3-amino-N-methoxy-N-methyl-5-nitro-benzamide (1(b)) (0.122 g) and EEDQ(0.201 g) in dichloromethane (20 ml) was stirred at ambient temperaturefor 16 hours. The solution was then stirred with 0.3M hydrochloric acid(20 ml) for ten minutes. The organic phase was separated, dried overmagnesium sulphate and evaporated under reduced pressure to give a gum.This was purified by chromatography using 1.ethyl acetate/hexane (50:50)2.ethyl acetate/hexane (75:25) to give the desired product (1) as acolourless gum (0.132 g).

NMR Spectrum (CDCl₃) δ2.35 (s, 3H), 2.62 (m, 2H), 3.4 (s, 3H), 3.44 (m,1H), 3.6 (s, 3H), 4.1 (m, 2H), 4.59 (t, 1H), 5.3 (m, 2H), 7.55 (d, 2H),8.09 (m, 1H), 8.25 (d, 2H), 8.3 (m, 1H), 8.6 (m,1H), 9.55 (br, s, 1H).

Starting material (1(c)) was synthesised as described in ReferenceExample 1-4 in European patent no 126587 (Sumitomo).

Starting material (1(b)) was prepared as follows. A mixture of3-amino-5-nitrobenzoic acid (10 g), pentafluoro-phenol (10 g) and DCCI(11.3 g) was stirred at ambient temperature for 24 hours. The reactionmixture was filtered and the filtrate poured onto a chromatographycolumn which was then eluted with ethyl acetate/hexane (10:90) to give3-amino-5-nitrobenzoic acid 2,3,4,5,6-pentafluorophenyl ester (1(a)) asa yellow solid (5.8 g).

NMR Spectrum (CDCl₃) δ4.3 (br, s, 2H), 7.7 (tr, 1H), 7.8 (tr, 1H), 8.36(tr, 1H).

A mixture of (1(a)) (1.0 g), N,O-dimethylhydroxylamine HCl salt (0.84 g)and triethylamine (1.82 ml) in dichloromethane (50 ml) was stirred atambient temperature for 48 hours. Water(50 ml) was added and the mixturestirred for a further 5 minutes. The organic phase was separated, driedover magnesium sulphate and evaporated under reduced pressure to give agum. This was purified by chromatography using 1. ethyl acetate/hexane(10:90), 2. ethyl acetate/hexane (50:50) as eluents to give startingmaterial 3-amino-N-methoxy-N-methyl-5-nitro benzamide (1(b)) as a yellowsolid (0.55 g).

NMR Spectrum: (CDCl₃) δ3.36 (s, 3H), 3.58 (s, 3H), 7.26 (tr, 1H), 7.56(tr, 1H), 7.90 (tr, 1H).

EXAMPLE 2

(See Scheme 2)

(2S,4S)-4-acetylsulfanyl-2[3-(N-methoxy-N-methylcarbamoyl)-5-nitro-phenylcarbamoyl]-pyrrolidine-1-carboxylicacid allyl ester

A mixture of (2S,4S),4-acetylsulfanyl-pyrrolidine-1,2-dicarboxylic acid1-allyl ester (1(d)) (0.2 g), 1(b) (0.165 g), and EEDQ (0.271 g), indichloromethane (20 ml) was stirred at ambient temperature for 16 hours.The solution was then stirred with 0.3M hydrochloric acid for a further10 minutes. The organic phase was then separated, dried over magnesiumsulphate and evaporated under reduced pressure. The product obtained waspurified by column chromatography using ethyl acetate/hexane (50:50) aseluent to give the desired product (2) as a colourless gum (0.152 g).

NMR Spectrum (CDCl₃) δ2.33 (s, 3H), 2.62 (m, 2H), 3.38 (m, 1H), 3.4 (s,3H), 3.6 (s, 3H), 4.05 (m, 2H), 4.59 (tr, 1H), 4.69 (d, 2H), 5.3 (m,2H), 5.95 (m, 1H), 8.14 (t, 1H), 8.28 (tr, 1H), 8.6 (tr, 1H), 9.7 (br,s,1H).

Synthesis of starting material (1(d)) is described as “Compound (A)” onpage 31 of International Patent Application No. WO 92/17479 (ImperialChemical Industries). Synthesis of starting material (1(b)) is describedin Example 1.

EXAMPLE 3

(See Scheme 3)

5-{[(2S,4S),4-acetylsulfanyl-1-(4-nitrobenzyloxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester

DMF (0.07 ml) was added to a stirred solution of oxalyl chloride (0.078ml) in dichloromethane (20 ml) cooled to −20° under an argon atmosphere.After 15 minutes a solution of (1(c)) (0.3 g; see Example 1) indichloromethane was added followed by a solution of N-methylmorpholine(0.099 ml) in dichloromethane (2 ml). After a further 15 minutes asolution of 5-amino-isophthalic acid allyl ester methyl ester (3(b))(0.192 g) in dichloromethane (5 ml) was added again followed by asolution of N-methylmorpholine (0.099 ml) in dichloromethane (2 ml). Themixture was allowed to warm to ambient temperature and stirred for 16hours. The reaction mixture was poured onto a flash column and elutedwith 1. ethyl acetate/hexane (50:50) and, 2. ethyl acetate/hexane(75:25) to give the desired end product (3) as a colourless gum (0.24g).

NMR Spectrum (CDCl₃) δ2.33 (s, 3H ), 2.62 (m, 2H), 3.45 (m, 1H), 3.95(s, 3H), 4.03 (m, 1H), 4.17 (m, 1H), 4.57 (tr, 1H), 4.85 (m, 2H), 5.32(m, 2H), 5.36 (m, 2H), 6.05 (m, 1H), 7.51 (m, 2H), 8.20 (m, 2H), 8.32(m, 2H), 8.34 (s, 1H), 9.2 (br, s, 1H).

Starting material (3(b)) was synthesised as follows. A mixture ofmono-methyl-5-nitroisophthalate (13.8 g), allyl bromide (7.96 g),potassium carbonate (13.94 g) and DMF (160 ml) was stirred at ambienttemperature for 4.5 h. The solid was filtered and DMF was evaporatedaway from the filtrate under reduced pressure. The residue was dissolvedin diethyl ether (300 ml) and water (100 ml) and stirred for fiveminutes. The organic layer was separated and washed with saturatedsodium bicarbonate solution (220 ml), brine (200 ml), dried overmagnesium sulphate and evaporated under reduced pressure to give5-nitro-isophthalic acid allyl ester methyl ester (3(a)) as a yellow oil(14.74 g).

NMR spectrum (CDCl₃) δ4.0 (s, 3H), 4.9 (m, 2H), 5.4 (m, 2H), 6.1 (m,1H), 9.0 (m, 3H).

A mixture of (3(a)) (15.46 g), tin (II) chloride dihydrate (65.78 g) andmethanol (200 ml) was stirred at reflux for 4 hours. Methanol wasevaporated under reduced pressure and the residue redissolved in ethylacetate (400 ml). Ammonia solution (sp. g. 0.880) was added dropwiseuntil the mixture reached pH 8 and no more precipitate was being formed.The solid was then filtered and the filtrate was washed with water (100ml), brine(100 ml), dried over magnesium sulphate and evaporated underreduced pressure to give starting material 3(b) as a yellow solid (13.56g).

NMR spectrum (CDCl₃) δ3.91 (s, 3H), 3.94 (s ,2H), 4.82 (m, 2H), 5.35 (m,2H), 6.05 (m, 1H), 7.52 (m, 2H), 8.08 (m, 1H).

EXAMPLE 4

(See Scheme 4)

5-{[(2S,4S),4-acetylsulfanyl-1-(carbamoylmethyl)-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester

A mixture of5-{[(2S,4S),4-acetylsulfanyl-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester TFA salt (4(e)) (0.12 g),iodoacetamide (0.085 g), sodium bicarbonate (0.058 g) and DMF (3.0 ml)was stirred at ambient temperature for 16 h. The DMF was evaporatedunder reduced pressure and the residue purified by chromatographyusing 1. ethyl acetate/hexane (60:40), 2. ethyl acetate and, 3.methanol/ethyl acetate (5:95) as eluents to give the desired product 4as a yellow solid (0.055 g).

NMR spectrum δ2.19 (2 tr,1H), 2.29 (s, 3H), 2.82 (m, 1H), 3.22 (m, 2H),3.48 (q, 2H), 3.6 (m, 1H), 3.94 (s, 3H), 4.05 (m, 1H), 4.85 (m, 2H),5.35 (m, 2H), 6.04 (m,1H), 6.1 (br, s, 1H), 6.30 (br, s, 1H), 8.43 (m,1H), 8.55 (m, 1H), 10.46 (br, s, 1H).

Starting material 4(e) was prepared as follows. A mixture of(2S,4S),4hydroxy-pyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester(1.0 g), EEDQ (1.6 g), Compound (3(b)) (see Example 3) anddichloromethane (100 ml) was stirred at ambient temperature for 16hours.

The mixture was poured onto a flash column and eluted with 1, ethylacetate/hexane (80:20) and, 2, ethyl acetate to give5-{[(2S,4S),4-hydroxy-1-(tert-butoxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester (4(a)) as a colourless gum (0.85 g.).

NMR Spectrum (DMSOd6) δ1.34 (2s, 9H), 1.97 (m, 1H), 2.15 (m, 1H), 3.30(m, 1H) 3.46 (m, 1H), 3.9 (s, 3H), 4.32 (m, 2H), 4.84 (d, 2H), 5.06 (d,1H), 5.35 (m, 2H), 6.07 (m, 1H), 8.18 (m, 1H), 854 (m, 2H).

A mixture of (4(a)) (0.8 g), methanesulphonyl chloride (0.152 ml),triethylamine (0.256 ml), and dichloromethane (20 ml) was stirred at 5°under an argon atmosphere for 10 minutes and then at ambient temperaturefor 2 h. Water (20 ml) was then added and the mixture stirred foranother 5 minutes. The organic phase-was separated, dried over magnesiumsulphate and evaporated under reduced pressure. The product was purifiedby chromatography using 1, ethyl acetate/hexane (30:70) and, 2, ethylacetate/hexane (80:20) as eluents to give5-{[(2S,4S),4methanesulfanyloxy-1-(tert-butoxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester (4(b)) as a clear oil (0.8 g).

NMR spectrum (CDCl₃) δ1.5 (s, 9H), 2.4 (m, 1H), 2.92 (m, 1H), 3.07 (s,3H), 3.63 (m, 1H), 3.9 (m, 1H), 3.95 (s, 3H), 4.66 (m, 1H), 4.85 (m,2H), 5.27 (m, 1H), 5.36 (m, 2H), 6.05 (m, 1H), 8.37 (m, 3H), 9.64 (br,s, 1H).

A mixture of 4(b) (0.74 g), potassium thioacetate (0.32 g) and acetone(25 ml) was maintained at reflux for 18 hours. The mixture was thencooled to room temperature and acetone evaporated under reducedpressure. The residue was dissolved in a mixture of ethyl acetate (50ml), 1.5M hydrochloric acid (25 ml), and ice (25 ml). The organic phasewas separated, dried over magnesium sulphate and evaporated underreduced pressure to give a red gum. This was purified by chromatographyusing 1, ethyl acetate/hexane (30:70) and, 2, ethyl acetate/hexane(70:30) to give5-{[(2S,4S),4-acetylsulfanyl-1-(tert-butoxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester (4(c)) as an orange gum (0.48 g).

NMR spectrum (CDCl₃) δ1.5 (s, 9H), 2.32 (s, 3H), 2.56 (m, 2H), 3.33 (m,1H), 3.93 (s, 3H), 4.04 (m, 2H), 4.52 (tr, 1H), 4.85 (m, 2H), 5.35 (m,2H), 6.05 (m, 1H), 8.38 (m, 3H), 9.63 (br, s, 1H).

A mixture of (4(c)) (3.6 g) and TFA (80 ml) was stirred at ambienttemperature for 10 minutes. TFA was evaporated under reduced pressureand the residue dissolved in ethyl acetate (200 ml.) and saturatedsodium bicarbonate solution (100 ml). This was then stirred for 10minutes, the organic phase separated, washed with water (100 ml) andbrine (100 ml) and dried over magnesium sulphate. The ethyl acetate wasremoved under reduced pressure and the residue purified bychromatography using 1, ethyl acetate/hexane (30:70), 2, ethylacetate/hexane (80:20) as eluents to give 4(f) (the free base which isused in Example 6) as a brown oil (2.3 g). NMR Spectrum (CDCl₃) δ2.05(m, 1H), 2.30 (s, 3H), 2.42 (br, s, 1H), 2.78 (m, 2H), 3.58 (m, 1H),3.85 (m, 1H), 3.94 (s, 3H), 3.99 (m, 1H), 4.84 (m, 2H), 5.35 (m, 2H),6.05 (m, 1H), 8.47 (m, 3H), 9.83 (br, s, 1H).

A mixture of (4(c)) (0.45 g) and TFA (10 ml) was stirred at ambienttemperature for 10 minutes. The TFA was evaporated away under reducedpressure and the residue purified by column chromatography using 1 ethylacetate/hexane (30:70), 2 ethyl acetate/hexane (60:40), 3 ethyl acetateand, 4 methanol/ethyl acetate (10:90) as eluents to give the desiredstarting material (4(e)) as a brown gum (0.46 g).

NMR Spectrum (CDCl₃) δ2.15 (m, 1H), 2.33 (s, 3H), 2.97 (m, 1H), 3.44 (m,1H), 3.91 (s, 3H), 3.97 (m, 1H), 4.08 (m, 1H), 4.82 (d, 2H), 4.98 (tr,1H), 5.35 (m, 2H), 6.03 (m, 1H), 8.12 (m, 2H), 8.26 (m, 1H).

EXAMPLE 5

(See Scheme 5)

5-{[(2S,4S),4-acetylsulfanyl-1-acetyl-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester

A mixture of (4(e)) (0.08 g; see Example 4), triethylamine (0.083 ml),acetic anhydride (0.056 ml) and dichloromethane (5 ml) was maintained atreflux for 16 hours. The mixture was cooled, evaporated under reducedpressure and purified by chromagraphy using 1 ethyl acetate/hexane(70:30), 2 ethyl acetate and, 3 methanol/dichloromethane (5:95) to givethe desired product 5 as a colurless gum (0.048 g).

NMR Spectrum (CDCl₃) δ2.18 (s, 3H), 2.35 (s, 3H), 2.48 (m, 1H), 2.77 (m,1H), 3.42(m, 1H),3.95 (s, 3H), 4.1 (m, 2H), 4.85 (m, 3H), 5.35 (m, 2H),6.06 (m, 1H), 8.40 (m, 3H), 9.88 (br, s, 1H).

Starting material 4(e) was prepared as described in Example 4.

EXAMPLE 6

(See Scheme 6)

5-{[(2S,4S),4-acetylsulfanyl-1-phenyloxycarbonyl-pyrrolidine-2-carbonyl]-amino}-isophthalicacid 1-allyl ester 3-methyl ester

A mixture of (4(f)) (0.07 g), phenyl chloroformate (0.026 ml),triethylamine (0.07 ml) and dichloromethane (3 ml) was stirred atambient temperature for 16 hours. The mixture was then evaporated underreduced pressure to give a gum which was purified by chromatographyusing 1 dichloromethane, 2 ethyl acetate/hexane (30:70) and, 3 ethylacetate/hexane (60:40) to give the desired product as a colourless gum(0.048 g.).

NMR Spectrum (DMSOd6) δ1.93-2.24 (m, 1H), 2.38 (s, H), 2.70 (m, 1H),3.63 (m, 1H), 3.91 (d, 3H), 4.18 (m, 2H), 4.60 (m, 1H), 4.87 (tr, 2H),5.38 (m, 1H), 6.08 (m, 1H), 6.70-7.69 (m, 5H), 8.20-8.53 (m, 3H), 10.61(d, 1H).

Starting material (4(f)) was prepared as described in Example 4.

EXAMPLE 7

(See Scheme 7)

5{[(2S,4S),4-acetylsulfanyl-1-(4-nitro-benzyloxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-3(N-methyl-methoxycarbamoyl)-benzoicacid allyl ester

A mixture of (1(c)) (0.02 g; see Example 1).3-amino-5(N-methyl-methoxycarbamoyl)-benzoic acid allyl ester (7(d))(0.16 g.), EEDQ (0.25 g) and dichloromethane (20 ml) was stirred for 16h at ambient temperature. The mixture was then washed with 0.3Mhydrochloric acid (30 ml), the organic phase separated. dried overmagnesium sulphate and evaporated to dryness under reduced pressure. Theresidue was purified by column chromatography using ethyl acetate/hexane(75:25) as eluent to give the desired product 7 as a yellow solid (0.053g).

NMR Spectrum (CDCl₃) δ2.33 (s, 3H), 2.60 (m, 2H), 3.38 (s, 3H), 3.42 (m,1H), 3.60 (s, 3H), 4.04 (m, 1H), 4.15 (m, 1H), 4.55 (m, 1H), 4.83 (m,2H), 5.30 (m, 2H), 5.35 (m, 2H), 6.04 (m, 1H), 7.52 (m, 2H), 8.10 (m,3H), 8.18 (m, 2H), 9.12 (br, s, 1H).

Starting material (1(c)) was prepared as described in Example 1.Starting material 7(d) was prepared as follows. A mixture of potassiumcarbonate (17.00 g), 5-nitroisophthalic acid (52.00 g), allyl bromideand dimethylacetamide (400 ml) was stirred at 90° for 4 h.Dimethylacetamide was evaporated away under reduced pressure and theresidue was dissolved in ethyl acetate. washed with water (2×300 ml) andthen extracted with aqueous saturated sodium bicarbonate solution (3×300ml). The extracts were combined. acidified to pH 4 with concentratedhydrochloric acid and reextracted with ethyl acetate (2×300 ml). Theextracts were combined, washed with water (300 ml), dried over magnesiumsulphate and evaporated under reduced pressure to give5-nitro-isophthalic acid 3-allyl ester (7(a)) as a cream solid (39.48g).

NMR Spectrum (CDCl₃/DMSOd6) δ4.90 (m, 2H), 5.42 (m, 2H), 6.08 (m, 1H),9.00 (m, 3H).

A solution of 7(a) (10.00 g), N-hydroxysuccinimide (5.04 g) and DCCI(9.03 g) in dichloromethane(400 ml) was stirred at ambient temperaturefor 3.5 h. The white precipitate which formed was filtered off and thefiltrate evaporated under reduced pressure to give a yellow oil. Thiswas purified by flash chromatography eluting with ethyl acetate/hexane(75:25) to give 5-nitro-isophthalic acid 1-(2,5-dioxo-pyrrolidin-1-yl)ester 3-allyl ester (7(b)) as a yellow solid (7.58 g).

NMR Spectrum (CDCl₃) δ2.95 (s, 4H), 4.92 (m, 2H), 5.43 (m, 2H), 6.07 (m,1H), 9.12 (m, 3H).

A mixture of (7(b)) (2.00 g), N,O-dimethylhydroxylamine hydrochloride(0.62 g), triethylamine (0.86 ml) and dichloromethane (60 ml) wasstirred at 5° for 30 min and then allowed to warm to ambient temperatureand stirred for a further 16 h. The mixture was poured onto a flashcolumn and eluted with ethyl acetate/hexane (40:60)to give3-(N-methyl-methoxycarbamoyl)-5-nitro benzoic acid allyl ester (7(c)) asa yellow oil.

NMR Spectrum (CDCl₃) δ3.43 (s, 3H), 3.58 (s, 3H), 4.90 (m, 2H), 5.40 (m,2H), 6.07 (m, 1H), 8.71 (m, 1H), 8.76 (m, 1H), 8.95 (m, 1H).

A mixture of (7(c)) (1.11 g), tin(II) chloride dihydrate (4.26 g) andmethanol (60 ml) was heated under reflux for 1 hour. The reactionmixture was cooled and the methanol evaporated away under reducedpressure. The residue was redissolved in ethyl acetate (100 ml) andammonia solution (sp. g. 0.880) was added dropwise until the solutionreached pH 8. The precipitate that formed was filtered and washed withethyl acetate (2×100 ml). The combined fitrate and washings wereevaporated under reduced pressure to give the desired starting material3-amino-5-(N-methyl-methoxycarbamoyl)-benzoic acid allyl ester (7(d)) asa white solid (0.610 g).

NMR Spectrum (CDCl₃) δ3.35 (s, 3H), 3.59 (s, 3H), 3.90 (br, s, 2H), 4.82(m, 2H), 5.35 (m, 2H), 6.04 (m, 1H), 7.15 (m, 1H), 7.45 (m, 1), 7.72 (m,1H).

EXAMPLE 8

(See Scheme 8)

5{[(2S,4S),4-acetylsulfanyl-1-4-nitro-benzyloxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-3(N-methyl-allyloxycarbamoyl)benzoicacid allyl ester

A mixture of (1(c)) (0.293 g; see Example 1),3-amino-5(N-methyl-allyloxycarbamoyl)-benzoic acid allyl ester (8(c))(0.210 g). EEDQ (0.268 g) and dichloromethane (20 ml) was stirred atambient temperature for 16 hours. The mixture was then washed with 0.3Mhydrochloric acid (30 ml), dried over magnesium sulphate and placedstraight onto a flash column eluting with ethyl acetate/hexane (75:25).The product obtained was placed onto a flash column eluting withmethanol/dichloromethane (2.5:97.5) to give the desired product 8 as aclear gum (0.153 g).

NMR Spectrum (CDCl₃) δ2.33 (s, 3H), 2.61 (m, 2H), 3.40 (s, 3H), 3.42 (m,1H), 4.04 (m, 1H), 4.15 (m, 1H), 4.26 (d, 2H), 4.55 (m, 1H), 4.83 (m,2H), 5.30 (m, 6H), 5.75 (m, 1H), 6.04 (m, 1H), 7.53 (m, 2H), 8.12 (m,2H), 8.21 (m, 3H), 9.12 (br, s, 1H).

Starting material (8(c)) was prepared as follows. A mixture of 7(b)(2.00 g; see Example 7), N-methylhydroxylamine hydrochloride (1.06 g)triethylamine (1.72 ml) and dichloromethane (60 ml.) was stirred at 5°or 30 minutes. It was then allowed to warm to ambient temperature andstirred for a further 16 hours. The reaction mixture was then poureddirectly onto a flash column eluting with ethyl acetate/hexane (50:50)to give 3-(N-methyl-hydroxycarbamoyl)-5-nitro-benzoic acid allyl ester(8(a)) as a cream solid (1.43 g).

NMR Spectrum (CDCl₃) δ3.48 (s, 3H), 4.90 (m, 2H), 5.42 (m, 2H), 6.05 (m,1H), 8.28 (br, s, 1H), 8.55 (m, 1H), 8.63 (m, 1H ), 8.96 (m, 1H).

A mixture of (8(a)) (0.60 g), allyl bromide (0.28 g), potassiumcarbonate (0.59 g) and DMF (20 ml) was stirred for 3 hours at ambienttemperature under an argon atmosphere. The dimethyl formamide was thenevaporated under reduced pressure and the residue dissolved in ethylacetate (50 ml) and water (50 ml). The organic phase was separated,washed with brine (50 ml), dried over magnesium sulphate and evaporatedunder reduced pressure to dryness to give3-(N-methyl-allyloxycarbamoyl)-5-nitro-benzoic acid allyl ester (8(b))as a yellow oil (0.571 g).

NMR Spectrum δ3.47 (s, 3H), 4.25 (m, 2H), 4.90 (m, 2H), 5.35 (m, 4H),5.65 (m, 1H), 6.06 (m, 1H), 8.73 (m, 1H), 8.78 (m, 1H), 8.95 (m, 1H).

A mixture of (8(b)) (0.523 g), tin(II) chloride dihydrate (1.84 g) andethyl acetate (50 ml) was heated under reflux for 6 hours. The mixturewas allowed to cool to ambient temperature and ammonia solution (sp. g.0.880) was added dropwise until the solution reached pH 8. The whiteprecipitate which had formed was filtered off, washed with ethyl acetate(2×50 ml) and the combined washings and filtrate evaporated to drynessto give the desired starting material (8(c)) as a yellow oil (0.472 g).

NMR Spectrum (CDCl₃) δ3.38 (s, 3H), 3.88 (m, 2H), 4.25 (d, 2H), 4.80 (m,2H), 5.32 (m, 4H), 5.75 (m, 1H), 6.03 (m, 1H), 7.15 (m, 1H), 7.45 (m,1H), 7.75 (m, 1H).

EXAMPLE 9

(see Scheme 9)

5-{[(2S,4S),1-(allyloxycarbonyl)-4-sulfonyl-pyrrolidine-2-carbonyl]-amino}-3(N-methyl-allyloxycarbamoyl)-benzoicacid allyl ester

An aqueous solution of 0.1M sodium hydroxide (4.41 ml) was added to asolution of5-{[(2S,4S),4-acetylsulfanyl-1-(allyloxycarbonyl)-pyrrolidine-2-carbonyl]-amino}-3(N-methyl-allyloxycarbamoyl)-benzoicacid allyl ester (9(a)) in allyl alcohol (15 ml) and the mixture wasthen stirred at ambient temperature for 1 hour. Hydrochloric acid (1.5M)was then added to bring the solution to pH3 and it was then evaporatedto dryness under reduced pressure. The residue was dissolved in ethylacetate (40 ml) and washed with water (2×40 ml). The organic phase wasseparated, dried over magnesium sulphate and evaporated to dryness togive a yellow foam. This was purified by chromatography using ethylacetate/hexane (75:25) as eluent to give the desired product 9 as ayellow gum (0.148 g).

NMR Spectrum (CDCl₃) δ1.88 (d, 2H), 2.62 (m, 2H), 3.37 (s, 3H), 3.45 (m,2H), 3.60 (s, 3H), 4.08 (m, 1H), 4.52 (tr, 1H), 4.65 (m, 2H), 4.83 (m,2H), 5.35 (m, 4H), 6.00 (m, 2H), 8.10 (m, 1H), 8.15 (m, 1H), 8.21 (m,1H), 9.15 (br, s, 1H).

Starting material 9(a) was prepared as follows. A mixture of 7(d) (0.568g; see Example 7), 1(d) (0.645 g; see Example 20), EEDQ (0.585 g) anddichloromethane (50 ml) was stirred at ambient temperature for 16 hours.The mixture was then washed with 0.3M hydrochloric acid(50 ml), driedover magnesium sulphate and applied to a flash column eluting with ethylacetate/hexane (75:25). It was further purified with a second columneluting with ethyl acetate/hexane (50:50) to give the desired startingmaterial (9(a)) as a colourless gum (0.401 g).

NMR Spectrum (CDCl₃) δ2.33 (s, 3H), 2.60 (m, 2H), 3.37 (s, 3H), 3.40 (m,1H), 3.61 (s, 3H), 4.02 (m, 1H), 4.13 (m, 1H), 4.58 (tr, 1H), 4.68 (m,2H), 4.83 (m, 2H), 5.35 (m, 4H), 6.00 (m, 2H), 8.10 (m, 1H), 8.14 (m,1H), 8.22 (m, 1H), 9.30 (br, s, 1H).

EXAMPLE 10

(see Scheme 10)

5-[((2S,4S),1-ayllyoxycarbonyl-4-sulfonyl-pyrrolidin-2-yl-methyl)-carbamoyl]-pyridine-2-carboxylicacid methyl ester

To a stirring solution of5-[((2S,4S),1-allyloxycarbonyl-4-BOCsulfanyl-pyrrolidin-2-yl-methyl)-carbamoyl]-pyridine-2-carboxylicacid methyl ester (10(a)) (991 mg; 2.07 mmole) in dichloromethane. TFA(6 mL; 78 mmole) was added dropwise. The solution was stirred, underargon, for 4 hours. The solvent and excess TFA were removed in vacuo.The residue was azeotroped with toluene (2×10 mL). Keeping exposure toair to a minimum the resultant oil was triturated with diethyl ether (20mL). The resultant solid was washed with cold diethyl ether (10 mL) anddried under high vacuum yielding the desired product 10 as a creamsolid, 654 mg (76%).

[4] has NMR (CDCl₃; 250 MHz) d 1.70 (m, 1H), 1.75 (d, 1H), 2.63-2.77 (m,1H), 3.15-3.50 (m, 3H), 3.90-4.00 (m, 1H), 4.05 (s, 3H), 4.07-4.23 (m,2H), 4.63 (m, 2H), 5.23-5.37 (m, 2H), 5.85-6.03 (m, 1H), 8.22 (d, 1H),8.35 (dd, 1H), 8.95 (s(br), 1H), 9.20 (s, 1H).

MS (FAB) m/z 380 (M+H)⁺.

Anal. C₁₇H₂₁N₃O₅S. 0.33 C₂HF₃O₂ 417: C, 50.9 (50.8); H, 5.3 (5.1); N,10.1 (10.1).

Starting material (10(c)) was prepared as follows. Pyridine2,5-dicarboxylic acid 2-methyl ester (10(a)) (9.0 g; 0.05 mole) wasadded to stirring thionyl chloride (25 mL) and the mixture refluxedgently for 2.5 hours. The excess thionyl chloride was removed in vacuoand the residual solid azeotroped with toluene (2×25 mL) to give5-chlorocarbonyl-pyridine-2-carboxylic acid methyl ester (10(b)) whichwas used crude in the next reaction.

To a stirring solution of compound (15(b)) (Example 15)(220 mg; 0.7mmole) in acetonitrile (6 mL) was added a solution of (10(b)) (0.7mmole) in acetonitrile (4 mL). Triethylamine (0.29 mL; 2.1 mmole) wasadded and the solution stirred for 23 hours. The solvent and excesstriethylamine were removed in vacuo and the residue partitioned betweenchloroform and water. The organic phase was washed with water, aqueoussodiun hydrogen carbonate solution and brine, dried over magnesiumsulphate and taken to dryness. The residual orange gum was flashchromatographed on kieselgel 9385, eluting initially with iso-hexanethen with increasing proportions of ethyl acetate. The desired startingmaterial 10(c) was isolated as a white foam (200 mg; 60%).

NMR (CDCl₃; 250 MHz) 1.50 (s, 9H), 1.80 (m, 1H), 2.62-2.75 (m, 1H),3.30-3.37 (m, 1H), 3.39-3.50 (m, 1H), 3.68-3.80 (m, 1H), 3.83-3.95 (m,1H), 4.03 (s, 3H), 4.13-4.28 (m, 2H), 4.62 (m, 2H), 5.20-5.37 (m, 2H),5.87-6.02 (m, 1H), 8.2 (d, 1H), 8.3 (dd, 1H), 8.87 (s, 1H), 9.2 (s, 1H).

MS (FAB) m/z 480 (M+H).

Anal. C₂₂H₂₉N₃O₇S 479: C, 55.1 (55.1); H, 6.4 (6.1); N, 8.5 (8.8).

EXAMPLE 11

(see Scheme 11)

(2S,4S)2-{[(5-ethoxycarbonyl-thiophene-2-carbonyl)-amino]-methyl}-4-sulfonyl-pyrolidine-1-carboxylicacid allyl ester

TFA (2 mL; 26 mmole) was added to a stirring solution of(2S,4S)2-{[(5-ethoxycarbonyl-thiophene-2-carbonyl)-amino]-methyl}-4-BOCsulfanyl-pyrollidine-1-carboxylicacid allyl ester (11(b)) (130 mg; 0.26 mmole) in dichloromethane (20mL). The solution was stirred under argon for 19 hours. The solvent andexcess TFA were removed in vacuo and the residue dried under high vacuumto give the desired product 11 as a water-white gum (64%).

NMR (CDCl₃; 250 MHz) d 1.38 (t, 3H), 1.55-1.70 (m, 1H), 1.75(d, 1H),2.60-2.76 (m, 1H), 3.10-3.50 (m, 3H), 3.80-3.95 (m, 1H), 4.05-4.25 (m,2H), 4.38 (q, 2H), 4.70 (m, 2H), 5.20-5.40 (m, 2H), 5.85-6.05 (m, 1H),7.47 (d, 1H), 7.73 (d, 1H), 8.52 (s(br),1H)

MS (FAB) m/z 399 (M+H)⁺ Anal. C₁₇H₂₂N₂O₅S₂ 0.5 C₂HF₃ O₂ 455: C, 47.6(47.5); H, 5.2 (4.9); N, 6.1 (6.15).

Starting material 11(b) was prepared in an analogous manner to theequivalent step in Example 10 but with addition of5-chlorocarbonyl-thiophene-2-carboxylic-acid-ethyl-ester (11(a)) tocompound (15(b)) (Example 15) with similar chromatographic work up,11(b) is a tacky water white gum. Yield 60%. Preparation of (11(a)) isdescribed in Journal of the American Pharmaceutical Association (Sci.Ed.) Vol. 41 pp 273-276 (1952).

NMR of 11(b): (CDCl₃; 250 MHz) d 1.4 (t, 3H), 1.5 (s, 9H), 1.70-1.85 (m,1H), 2.57-2.73 (m, 1H), 3.26-3.36 (m, 1H), 3.38-3.50 (m, 1H), 3.65-3.87(m, 2H), 4.10-4.25 (m, 2H), 4.35 (q, 2H), 4.65 (m, 2H), 5.20-5.38 (m,2H), 5.85-6.04 (m, 1H), 7.47 (d, 1H), 7.72(d, 1H), 8.45 (s(br), 1H).

MS (FAB) m/z 499 (M+H)⁺, other m/z 183.

Anal. C₂₂H₃₀N₂O₇S₂ 498 C, 53.4 (53.0); H, 6.3 (6.1); N, 5.5 (5.6).s

EXAMPLE 12

(see Scheme 12)

N-(3,4-dichlorobenzyl)-N′-((2S,4-S),4-sulfonyl-pyrrolidin-2-yl-methyl)thiophene-2,5-dicarboxamide

To a stirring solution ofN-(3,4-dichlorobenzyl)-N′-((2S,4S),-1-allyloxycarbonyl-4-sulfonyl-pyrrolidin-2-yl-methyl)thiophene-2,5-dicarboxamide(12(e)) (59 mg; 0.1 mmole) in dichloromethane (10 mL), under argon, wasadded trimethylsilyliodide (0.35 mL; 0.25 mmole). After 20 hours atambient temperature the dichloromethane and excess trimethylsilyliodidewere removed in vacuo and the residue treated with methanol (3 mL). Theinsoluble material was treated with farther methanol (2×3 mL) and thentriturated with diethyl ether to yield a solid which was filtered anddried to give the desired product 12 as a light brown solid (59%).

NMR (DMSO-d₆; 250 MHz) δ1.65-1.90 (m, 1H), 2.50-2.62 (m, 1H), 3.20-3.40(m, 2H), 3.55-3.70 (m, 2H), 3.75-3.90 (m, 2H), 4.45 (d, 2H), 7.32 (m,1H), 7.58 (m, 2H), 7.73 (d, 1H), 7.78 (d, 1H), 8.68 (br, 1H), 8.88 (t,1H), 9.22 (t, 1H).

MS (FAB) m/z 444 (M+H)⁺ other 111, 312 Anal. C₁₈H₁₉Cl₂N₃O₂S₂ 1.25 HI 0.5C₄H₁₀O 640 C, 37.6 (37.5); H, 3.5 (3.9); N, 6.5 (6.6).

Starting material (12(e)) was prepared as follows. To a stirringsolution of 3,4-dichlorobenzylamine (0.53 mL; 4.0 mmole) in acetonitrile(10 mL) was added triethylamine (1.67 mL; 12.0 mmole) and a solution of(11(a)) (0.87 g; 4.0 mmole, see Example 11) in acetonitrile (20 mL). Thesolution was stirred at ambient temperature. under argon, for 22 hours.The solvent and excess triethylamine were removed in vacuo and theresidue partitioned between chloroform and water. The organic phase waswashed with water and brine, dried over magnesium sulphate and vacuumedto dryness to give5-(3,4-dichlorobenzyl-carbamoyl)-thiophene-2-carboxylic acid ethyl ester(12(a)) as a cream solid ( 90%).

NMR (CDCl₃; 250 MHz) δ1.40 (t, 3H), 4.38 (q, 2H), 4.57 (d, 2H), 6.47(t(br), 1H), 7.28 (m, 1H), 7.42 (m, 2H), 7.48 (d, 1H), 7.73 (d, 1H) MS(CI) m/z 358 (M+H)⁺

Anal. C₁₅H₁₃Cl₂NO₃S 358: C, 50.4 (50.3); H, 3.8 (3.7); N, 3.9 (3.9).

Aqueous 1M sodium hydroxide (16.3 mL; 16.3 mmole) was added to astirring, solution of (12(a)) (1.17g; 3.3 mmole) in ethanol (70 mL). Thereaction mixture was and adjusted to pH 2 by addition of 2M hydrochloricacid. The filtered solid was washed with water and dried in vacuo togive 5-(3,4-dichlorobenzyl-carbamoyl)-thiophene-2-carboxylic acid(12(b)) as a white solid (83%).

NMR (DMSO d6; 200 MHz) d 4.43 (d, 2H), 7.3 (dd, 1H), 7.58 (m, 2H), 7.68(d, 1H), 7.78 (d, 1H), 9.28 (t, 1H) MS (CI) m/z 330 (M+H)⁺

Anal. C₁₃H₉Cl₂NO₃S 330 C, 47.3 (47.3); H, 2.7 (2.7); N, 4.2 (4.2).

A stirring solution of (12(b)) (495 mg; 1.5 mmole) in dichloromethane(25 mL) was cooled in an ice bath and DMF (1 drop) and oxalyl chloride(0.175 mL; 2.0 mmole) added dropwise. The solution was stirred atambient temperature under argon for 4 hours. The dichloromethane andexcess oxalyl chloride were removed in vacuo. The residue was azeotropeduith toluene (2×15 mL) to give5-(3,4-dichlorobenzyl-carbamoyl)-thiophene-2-carbonyl-chloride (12(c))which was used crude in the next step.

Triethylamine (0.83 mL; 4.5 mmole) and a solution of compound (15(b))(Example 15) (316 mg; 1.0 mmole) in acetonitrile (10 mL) were added to astirring mixture of (12(c)) (1.5 mmole) in acetonitrile (15 mL) andstirred at ambient temperature under argon for 19 hours. Theacetonitrile and excess triethylamine were removed in vacuo and theresidue partitioned between chloroform and water. The organic phase waswashed with water and brine, dried over magnesium sulphate and vacuumedto dryness to giveN-(3,4-dichlorobenzyl)-N′-((2S,4S),-1-allyloxycarbonyl-4-BOCsulfanyl-pyrrolidin-2-yl-methyl)thiophene-2,5-dicarboxamide(12(d)) as a tacky brown solid (95%).

NMR (CDCl₃; 200 MHz) δ1.5 (s, 9H), 1.65-1.85 (m, 1H), 2.47-2.73 (m, 1H),3.25-3.50 (m, 2H), 3.65-3.85 (m, 2H), 4.10-4.23 (m, 2H), 4.57 (d, 2H),4.64 (m, 2H), 5.20-5.40(m, 2H), 5.85-6.05 (m, 1H), 6.45 (t, 1H), 7.20(dd, 1H), 7.40 (m, 2H), 7.46 (d, 1H), 7.53 (d, 1H), 8.47 (br, 1H) MS(FAB) m/z 628 (M+H )⁺ Anal. C₂₇H₃₁ Cl₂N₃O₆S .H₂O 646 C, 50.2 (50.2); H,4.9 (5.1); N, 6.5 (6.5).

TFA (5 mL; 65 mmole) was added to a stirred solution of (12(d)) (600 mg;0.93 mmole) in dichloromethane (25 mL). The solution was stirred atambient temperature under argon for 4 hours, solvent and excess TFA wereremoved in vacuo and the residue azeotroped % with toluene to give thedesired starting material (12(e)).

NMR (CDCl₃; 250 MHz) δ1.55-1.75 (m, 1H), 1.75 (d, 1H), 2.50-2.72 (m,1H), 3.12-3.43 (m, 1H), 3.65-3.90 (m, 2H), 4.03-4.20 (m, 2H), 4.54 (d,2H), 4.63 (m, 2H), 5.17-5.37 (m, 2H), 5.85-6.03 (m, 1H 0, 6.63 (br, 1H),7.10-7.55 (m, 5H), 8.5 (br, 2H)

MS (FAB) m/z 528 (M+H)⁺ Anal. C₂₂H₂₃Cl₂N₃O₄S₂ 0.33 C₄H₁₀O 0.3 C₂HF₃O₂586.5 C, 49.0 (49.0); H, 4.5 (4.6); N, 7.2 (7.2).

EXAMPLE 13

(see Scheme 13)

5-[N-(3,4-dichlorobenzyl)carbamoyl]-N-((2S,4S)-4-sulfanylpyrrolidin-2-yl-methyl)pyridine-2-carboxamide

5-[N-(3,4-dichlorobenzyl)carbamoyl]-N-((2S,4S)-1-allyloxycarbonyl-4-sulfanylpyrrolidin-2-yl-methyl)pyridine-2-carboxamide(13(e)) was treated with trimetiylsilyliodide in similar manner tocompound (12(e)) in Example 12. The desired product 13 was obtained as amedium brown solid (26%).

NMR (DMSO-d6; 200 MHz) δ1.70-1.82 (m, 1H), 3.15-3.40 (m, 2H), 3.55-3.90(m, ?H), 4.52 (d, 2H), 7.35 (dd, 1H), 7.60 (m, 2H), 8.18 (d, 1H), 8.47(dd, 1H), 8.75 (br, 1H), 9.10 (d, 1H), 9.28 (t+?, 2H), 9.42 (t, 1H).

MS (FAB) m/z 439 (M+H)⁺, Anal. C₁₉H₂₀Cl₂N₄O₂S. 1.5 HI.0.33 C₄H₁₀O 655.7C, 37.4 (37.2); H, 3.4 (3.7); N, 8.1 (8.5).

Starting material (13(e)) was prepared as follows,5-chlorocarbonyl-pyridine-2-carboxylic acid methyl ester was reactedwith 3,4-dichlorobenzylamine analogously with preparation of compound(12(a)) in Example 12 to obtain5(3,4-dichlorobenzylcarbamoyl)-pyridine-2-carboxylic acid methyl-ester(13(a)) as a cream solid (61%).

NMR (CDCl₃; 250 MHz), d 4.05 (s, 3H), 4.62 (d, 2H), 6.80 (t(br), 1H),7.22 (dd, 1H), 7.43 (m, 2H), 8.20 (d, 1H), 8.30 (m, 1H), 9.08 (d, 1H).MS (CI) m/z 339 (M+H)⁺

Anal. C₁₅H₁₂Cl₂N₂O₃ 339 C, 53.2 (53.1); H, 3.5 (3.6); N, 8.1 (8.3).

Compound (13(a)) was treated in an analogous manner to compound (12(a))in Example 12 to obtain5(3,4-dichlorobenzylcarbamoyl)-pyridine-2-carboxylic acid (13(b)) as anoff-white solid (82%).

NMR (DMSO-d₆; 200 MHz) δ4.50 (d, 2H), 7.33 (dd, 1H), 7.58 (m, 2H), 8.13(d, 1H), 8.37 (dd, 1H), 9.12 (d, 1H), 9.40 (t, 1H) MS (CI) m/z 325(M+H)⁺

Anal. C₁₄H₁₀Cl₂N₂O₃.H₂O 343 C, 48.9 (48.9); H, 3.5 (3.5); N, 8.0 (8.2).

Compound (13(b)) was treated in an analogous manner to compound (12(b))in Example 12 to give5(3,4-dichlorobenzylcarbamoyl)pyridine-2-carbonylchloride (13(c)) whichwas used crude in the next reaction.

Compound (13(c)) was reacted with compound (15(b)) (Example 15) in asimilar manner to compound (12(c)) in Example 12 to give5-[N-(3,4-dichlorobenzyl)carbamoyl]-N-((2S,4S)-1-allyloxycarbonyl-4-BOCsulfanylpyrrolidin-2-yl-methyl)pyridine-2-carboxamideas a light brown solid (13(d)) (81%).

NMR (CDCl₃; 250 MHz) δ1.50 (s, 9H), 1.73-1.90 (m, 1H), 2.50-2.65 (m,1H), 3.20-3.30 (m, 1H), 3.62-3.80 (m, 2H), 4.10-4.27 (m, 2H), 4.65 (d?,4H), 5.18-5.38 (m, 2H), 5.83-6.05 (m, 1H), 6.80 (t(br), 1H), 7.20-7.28(m, 2H), 7.40-7.48 (m, 2H), 8.23 (s, 2H), 8.75 (br, 1H), 8.98 (d?, 1H).

MS (FAB) m/z 623 (M+H)⁺ Anal. C₂₈H₃₂Cl₂N₄O₆S 623 C, 53.8 (53.9); H, 5.1(5.2); N 8.9 (9.0) mp 136-137.5° C.

Compound (13(d)) was treated in a similar manner to compound (12(d)) inExample 12 to give the desired starting material (13(e)) as a lightbrown solid (64%).

NMR (CDCl₃; 250 MHz) δ1.70 (d, 1H), 1.80-2.00 (m, 1H), 2.52-2.65 (m,1H), 3.05-3.25 (m, 2H), 3.60-3.85 (m, 2H), 4.05-4.20 (m, 2H), 4.60 (d?,4H), 5.18-5.33 (m, 2H), 5.85-6.03 (m, 1H), 6.80 (br, 1H), 7.20 (dd, 1H),7.40-7.47 (m, 2H), 8.23 (s, 2H), 8.78 (br, 1H), 9.0 (s, 1H), MS (FAB)m/z 523 (M+H)⁺

Anal. C₂₃H₂₄Cl₂N₄O₄S. 0.1 C₂HF₃O₂ 534.4 C, 52.4 (52.1); H, 4.6 (4.5); n10.3 (10.5) mp 101-105° C.

EXAMPLE 14

(see Scheme 14)

1-hydroxy-4-[((2S,4S),4-sulfonyl-pyrrolidin-2yl-methyl)-amino-sulfonyl]naphthalene-2-carboxylic-acid

To a stirring solution of1-hydroxy-4-[((2S,4S),1-allyloxycarbonyl-4-sulfonyl-pyrrolidin-2yl-methyl)-aminosulfonyl]-naphthalene-2-carboxylic-acid(14(c)) (47.5 mg; 0.1 mmole) in dichloromethane (10 mL) was added TMSI(0.56 mL; 0.4 mmole). The solvent and excess TMSI were removed in vacuoafter 6 hours. Methanol (5 mL) was added to the residue and then removedin vacuo from the solution. The residue was triturated with diethylether, filtered and dried in vacuo to obtain the desired product 14 as abrown solid (74%).

NMR (DMSO-d6; 250 MHz) δ1.45-1.62 (m, 1H), 2.25-2.45 (m, 1H), 2.90-3.25(m, 3H), 3.45-3.70 (m, 2H), 7.72 (m, 1H), 7.85 (m, 1H), 8.12 (m, 1H),8.38-8.60 (m, 2H), 9.15 (br, 1H)

MS (FAB) m/z 3.83 (M+H)⁺

Anal. C₁₆H₁₈N₂O₅S₂. 1.25 HI.0.5 C₄H₁₀O 579 C, 37.0 (37.3); H, 4.1 (4.2);N, 4.8 (4.8).

Starting material (14(c)) was prepared as follows. Compound (15(b))(Example 15) and 1-hydroxy-4-chlorosulfonyl-naphthalene-2-carboxylicacid (14(a)) were coupled in a similar manner to the equivalent step inExample 15 to give 1-hydroxy-4-[((2S,4S),1-allyloxycarbonyl-4-BOCsulfonyl-pyrrolidin-2yl-methyl)-aminosulfonyl]-naphthalene-2-carboxylic-acid(14(b)) as a light brown solid (80%).

NMR (CDCl₃; 250 MHz) δ1.45 (s, 9H), 1.50-1.75 (m, 1H), 2.28-2.42 (m,1H), 2.96-3.10 (m, 2H), 3.48-3.60 (m, 1H), 3.80-3.90 (m, 1H), 3.95-4.05(m, 1H), 4.47 (m, 2H), 4.53-4.63 (m, 1H), 7.55 (m, 1H), 7.67 (m, 1H),8.50 (m, 2H), 8.70 (m, 1H) MS (FAB) M+Na⁺ 589, other 317.261 Anal.C₂₅H₃₀N₂O₉S₂.H₂O.0.8 C₃H₁₅N 664.8 C, 53.7 (53.8); H, 6.7 (6.6); N, 5.9(5.9).

2M Aqueous sodium hydroxide (5 mL; 10.0 mmole) was added to a stirringsolution of (14(b)) (333 mg; 0.5 mmole) in methanol (5 mL). The solutionwas evaporated to dryness after 42 hours and the residue dissolved inwater (10 mL). The solution was adjusted to pH 2 with 2M hydrochloricacid and the solid was filtered, washed with water and dried in vacuo togive the desired starting material (14(c)) as a white solid (72%).

NMR (CDCl₃; 200 MHz) δ1.48-1.70 (m, 2H), 2.38-2.52(m, 1H), 2.85-3.40 (m,?H), 3.90-4.05 (m, 2H), 4.40-4.60 (m, 3H), 5.10-5.35 (m, 3H), 5.70-5.95(m, 2H), 6.20-6.45 (br, 1H), 7.57-7.90 (m, 3H), 8.43-8.70 (m, 4H)

MS (FAB) m/z 467 (M+H)⁺ Anal. C₂₀H₂₂N₂O₇S₂.0.5 H₂O 475 C, 50.6 (50.5);H, 4.8 (4.8); N, 6.0 (5.9).

EXAMPLE 15

(See Scheme 15)

(2S)-2-{3-[([2S,4S]-4-sulfanyl-pyrrolidin-2-yl-methyl)-sulfamoyl]-benzoylamino}-4-methylsulfanyl-butyricacid methyl ester

TFA (2.0 mL) was added to a stirred solution of(2S)-2-{3-[([2S,4S]-4-BOCsulfanyl-pyrrolidin-2-yl-methyl)-sulfamoyl]-benzoylamino}-4-methylsulfanyl-butyricacid methyl ester (15(d)) (101 mg, 0.18 mmol) in CH₂Cl₂ (2.0 mL) at roomtemperature under argon. After 1 h the reaction mixture was concentratedto a dryness, azeotroped with toluene (3×10 mL) and dried to yield thedesired product 15 as a colourless gum: 101.8 mg (99%).

¹H NMR (CDCl₃, 250 MH^(z)) δ1.6-1.8 (1H, m); 2.0 (1H, d, SH); 2.1-2.4(5H, m); 2.52.65 (3H, m); 3.15-3.4 (3H, m) 3.45-3.65 (1H, m); 3.7-3.85(4H, m) 3.9-4.1 (1H, m); 4.85-5.0 (1H, m); 7.55-7.7 (2H, m) 7.8 (1H, s);8.0 (1H, d); 8.1 (1H, d); 8.3 (1H, s); 9.0-9.4 (1H, s); 10.0-10.4 (1H,s).

MS (ESP+) m/z 462 (M+H)⁺.

Starting material (15(d)) was prepared as follows. Triethylamine (3.0mL, 21.5 mmol) was added to a stirred suspension of L-methionine methylester. HCl(4.37 g, 21.8 mmol) in CH₂Cl₂ (50 mL). The resulting mixturewas left to stir for 30 min at room temperature then filtered. Thefiltrates were then added to a stirred solution of3-chlorosulphonyl-benzoyl chloride (5.23 g, 21.9 mmol) and triethylamine(7.6 mL, 54.7 mmol) in CH₂Cl₂ (50 mL) at 0° under argon. The reactionmixture was allowed to warm to room temperature and quenched withice-water(100 mL). The organics were the dried over MgSO₄, filtered andconcentrated to a viscous brown gum. This was then purified by flashchromatography on 9385 SiO₂, eluting with 50% EtOAc/i-Hexane to give(2S)-2-(3-chlorosulfonyl-benzoylamino)-4-methylsulfonyl-butyric acidmethyl ester (15(a)) as a viscous orange oil; 2.88 g (36%).

¹H NMR (CDCl₃,250 MH_(Z)) δ2.1-2.2 (5H, m); 2.65 (2H, t); 3.83 (3H, s);4.95 (1H, m); 7.23 (1H, d); 7.74 (1H,t); 8.2 (2H,m); 8.47 (1H,m). MS(CI) m/z 366 (M+H)⁺, 332,300.

A solution of 15(a) (1.53 g, 4.18 mmol) in CH₂Cl₂ (20 mL) was added to astirred solution of(2S,4S)-2-aminomethyl-4-BOCsulfanyl-pyrollidine-1-carboxylic acid allylester (15(b)) (prepared as described in International Patent ApplicationWO 92/17480, see pages 39-41) (1.32 g, 4.18 mmol) and (^(i)Pr)₂NEt (1.5mL, 9.0 mmol) in CH₂Cl₂ (30 mL) at 0° C. under argon. The resultingsolution was allowed to warm to room temperature and stirred for 18hours. The reaction mixture was then washed with water (100 mL), driedover MgSO₄, filtered and concentrated to a viscous white gum. This wasthen purified by flash chromatography on 9385 SiO₂, eluting with agradient of 35-50% EtOAc/i-Hexane to give(2S,4S)-4-BOCsulfanyl-2-{[3-([1S]-1-methoxycarbonyl-3-propylcarbamoyl]-methyl}-pyrrolidine-1-carboxylicacid allyl ester (15(c)) as a colourless foam: 2.19 g (81.3%).

¹H NMR (CDCl₃,200 MH_(Z)) δ1.5 (9H, s); 1.65-1.9 (1H, s); 2.05-2.35 (5H,m); 2.4-2.7 (3H, m); 3.3-3.4 (3H, m); 3.55-3.75 (1H, m); 3.8 (3H, s);3.9-4.2 (2H, d); 4.98 (1H, m); 5.15-5.35 (2H, m); 5.8-6.0 (1H, m); 6.5(1H, s); 7.4 (1H, s); 7.55 (1H, t); 7.9-8.05 (2H, m); 8.25 (1H, m).

MS (FAB) m/z 646 (M+H)⁺, 590,568,546,230.

Anal. Calcd for C₂₇H₃₉N₃O₉S₃.0.3CH₂Cl₂:C, 48.8; H, 5.95; N, 6.26.

Found C, 48.9; H, 6.2; N, 6.0.

Tri-nButyl tin hydride (565 mL, 2.1 mmol) was added to a stirredsolution of (15(c)) (1.18 g, 1.8 mmol) and (PPh)₃PdCl₂ (13 mg, 0.018mmol) in a mixture of water (0.5 mL) and CH₂Cl₂ (100 mL). The reactionmixture was left to stir for 10 minutes, dried over MgSO₄, filtered andconcentrated to a brown oil. This was then purified by flashchromatography on 9385 SiO₂, eluting with a gradient of 0-10%EtOAc/i-Hexane to give the desired starting material 15(d) as a whitefoam: 751 mg (73%).

¹H NMR (CDCl₃+CD₃COOD,250 MH^(z)) δ1.5 (9H, s); 1.85-1.97 (1H, m);2.1-2.35 (5H, m); 2.45-2.7 (3H, m); 3.1-3.4 (3H, m); 3.65-4.25 (6H, m);4.9-5.0 (1H, m); 7.63 (1H, t); 7.97-8.05 (1H, m); 8.1-8.17 (1H, m)8.35-8.42 (1H, m).

MS (ESP+) m/z 562 (M+H)⁺, 462.

Anal. Calcd for C₂₃H₃₅N₃O₇S₃: C, 49.2; H, 6.28; N, 748.

Found C, 49.4; H, 6.3; N, 7.2.

EXAMPLE 16

(See Scheme 16)

(2S),2-{3-[([2S,4S]-4-sulfanyl-pyrrolidin-2-yl-methyl)-sulfamoyl]-benzoylamino}-4-methylsulfanyl-butyricacid

2N NaOH(2.0 mL, 4.0 mmol) was added to a stirred solution of compound(15(d)) (prepared in Example 15) (200 mg, 0.36 mmol) in MeOH at roomtemperature under argon. After 18 h the reaction mixture wasconcentrated to remove the MeOH. The resulting residues were dissolvedin H₂O (2.0 mL) and acidified to pH3 with 2N HCl. The resulting solutionwas purified by reverse phase HPLC (Dynamax C18.8μ prepcolunmn), elutingwith a gradient of 0-40% MeOH/H₂O. Product fractions were concentratedand azeotroped with toluene (3×25 mL) to give a colourless glass whichwas then triturated with Et₂O (25 mL), filtered and dried to yield thedesired product 16 as a white powder: 85.2 mg (54%).

¹H NMR (DMSO-D₆+CD₃COOD,250 MH^(Z)) δ1.45-1.65 (1H, m); 2.0-2.2 (5H, m);2.3-2.7 (3H+DMSO, m); 2.95-3.2 (3H, m); 3.35-4.2 (3H, m); 4.5-4.65 (1H,m); 7.65-7.8 (1H, m); 7.9-8.05 (1H, m); 8.1-8.25 (1H, m); 8.3-8.4 (1H,m).

MS (FAB) m/z 448 (M+H)⁺.

Anal. Calcd for C₁₇H₂₅N₃O₅S₃: C, 45.6; H, 5.63; N, 9.39.

Found C, 45.5; H, 5.8; N, 9.1.

EXAMPLE 17

(See Scheme 17)

N-(3,4-dichlorophenyl)-3-[([2S,4S],4-sulfanyl-pyrrolidin-2-yl-methyl)-sulfamoyl]-benzamide

N-(3,4-dichlorobenzyl)-3-[([2S,4S],4-BOCsulfanyl-pyrrolidin-2-yl-methyl)-sulfamoyl]-benzamide(17(c)) was deprotected with TFA (analogously to compound (15(d)) inExample 15) to give the desired product 17 in 97% yield aftertrituration with Et₂O.

¹H NMR (CDCl₃200 MH_(Z)) δ1.5-1.8 (1H, m); 1.8-2.2(2H+H₂O,m,SH,NH);2.5-2.7 (1H,m); 3.1-3.35 (3H, m); 3.4-4.1 (3H, m); 4.55(2H, d); 7.15 (1H, dd); 7.2 (1H, s); 7.32 (1H, d); 7.4 (1H, d); 7.65(1H+PPh₃PO, m); 7.9 (1H, m); 8.2 (1H, m); 8.2 (1H, m); 8.35 (1H, m);8.5-9.3 (1H, s, NH); 10.3-10.7 (1H, s, NH).

MS (ESP+) m/z 474 (M+H)⁺, 279(Ph₃PO)

Starting material (17(c)) was prepared as follows.3,4-Dichlorobenzylamine was coupled with 3-Chlorosulphonylbenzoylchloride (analogously as for compound (15(a)) in Example 15) to give3-(3,4-dichloro-benzylcarbamoyl)-benzene-sulfonyl-chloride (17(a)) in28% yield.

¹H NMR(CDCl₃,250 MH_(Z)) δ4.6 (2H, d); 6.6 (1H, s, NH); 7.2 (1H, dd),7.4-7.5 (2H, m); 7.75 (1H, t); 8.15-8.25 (2H, m) 8.4 (1H, m) MS (FAB)m/z 378 (M+H)⁺,380.

Compound 15(b) (Example 15) was coupled with (17(a)) analogously as forthe equivalent step in Example 15 to giveN-(3,4-dichlorobenzyl)-3-[([2S,4S],4-BOCsulfanyl-pyrrolidin-2-yl-methyl)-sulfamoyl]-benzamide (17(b)) in 72.5% yield.

¹H NMR (CDCl₃,200 MH_(Z)) δ1.5 (9H, s); 1.6-1.9 (1H+H₂O, m); 2.4-2.6(1H, m); 3.1-3.3 (3H, m); 3.6-3.7 (1H, m); 3.8-4.1 (2H, m); 4.4 (2H, d);4.6 (2H, d); 5.1-5.3 (2H, m); 5.7-5.95 (1H, m); 6.08 (1H, s, NH); 7.2(1H, dd); 7.35-7.7 (4H, m); 7.95 (1H, d); 8.15 (1H, d); 8.25-8.35 (1H,s, NH).

MS (FAB) m/z 658 (M+H)⁺

Anal. Calcd for C₂₈H₃₃N₃Cl₂O₇S₂: C, 51.1; H, 5.05; N, 6.38.

Found C, 50.8; H,5.2; N,6.2.

Compound (17(b)) was deprotected, analogously as for the equivalent stepin Example 15, to give the desired starting material (17(c)) in 70%yield.

¹H NMR (CDCl₃, 250 MH_(Z)) δ1.15-1.45 (1H, m); 1.5 (9H, s); 2.25-2.4(1H, m); 2.6-2.9 (4H, m); 3.02 (1H, dd): 3.25-3.4 (2H, m); 3.45-3.6 (1H,m); 4.6 (2H, m); 7.2 (1H, dd); 7.4 (1H, d); 7.45 (1H, d); 7.6 (1H, t);7.95 (1H, d); 8.1 (1H, d); 8.25 (1H, s).

MS (ESP+) m/z 574 (M+H)⁺,574,279 (PPh₃O)

EXAMPLE 18

(See Scheme 18)

N-(3,4-dichlorobenzyl)-N′-([2S,4S],4-sulfanyl-pyrrolidin-2yl-methyl)-isophthalamide

N-(3,4-dichlorobenzyl)-N′-([2S,4S],4-BOCsulfanyl-pyrrolidin-2yl-methyl)-isophthalamide(18(e)) was deprotected with TFA (analogously to the equivalent step inExample 15) to give the desired product 18 in 100% yield aftertrituration with Et₂O.

¹H NMR (CDCl₃+CD₃COOD, 250 MH_(Z)) δ1.75-1.9 (1H, m); 2.6-2.75 (1H, m);3.2-3.35 (1H, m); 3.45-3.65 (1H, m); 3.7-3.95 (3H, m); 4.05-4.15 (1H,m); 4.6 (2H, s); 7.2 (1H, dd); 7.4 (1H, d); 7.55 (1H, t); 7.95-8.05 (1H,m); 8.1-8.2 (1H, m); 8.4 (1H, m). MS (ESP+) m/z 438 (M+H)⁺.

Starting material (18(e)) was prepared as follows. A suspension ofisophthalic acid monomethyl ester (18(a)), (2.65 g, 14.7 mmol) in CH₂Cl₂(100 mL) and DMF (10 drops) was treated with oxallyl chloride (2.6 ml,29.8 mmol) at 0° under argon. The reaction mixture was allowed to warmto room temperature over 18 h. The resulting solution was concentratedand azeotroped with toluene to give a crystalline yellow solid. This wasthen redissolved in CH₂Cl₂ (100 mL) and added dropwise to a stirredsolution of 3,4-dichlorobenzylamine (2.6 g, 14.7 mmol) and Et₃N (5 mL,35.9 mmol) in CH₂Cl₂ (100 mL) at 0° under argon. The resulting solutionwas allowed to warm to room temperature over 4 hours, washed with 1NHCl(50 mL), saturated NaHCO₃ (aq) (50 mL), dried over MgSO₄, filteredand concentrated to an orange oil. This was then purified by flashchromatography on 9385 SiO₂ eluting on a gradient of 25-50%EtOAc/i-Hexane to yield 3-(3,4-dichlorobenzyl-carbamoyl)-benzoic acidmethyl ester (18(b)) as a pale yellow oil: 3.99 g (80%).

¹H NMR (CDCl₃,200 MH_(Z)) δ3.9 (3H, s); 4.6 (2H, d); 6.6-6.8 (1H, t,NH); 7.18 (1H, dd); 7.38-7.45 (2H, m); 7.54 (1H,t); 8.0-8.1 (1H, m);8.13-8.23 (1H, m); 8.35-8.42 (1H, m). MS (CI) m/z 338 (M+H)⁺.

A stirred solution of (18(b)) (3.85 g, 11.4 mmol) in MeOH (100 mL) atroom temperature under argon was treated with 2N NaOH (12 mL, 24 mmol).The reaction mixture was allowed to stir at room temperature for 4 h,concentrated to 1/5 volume and acidified to pH4 with 2N HCl. Theresulting precipitate was then collected by filtration, washed withwater (2×25 mL) and dried under high vacuum to yield3-(3,4-dichlorobenzyl-carbamoyl)-benzoic acid (18(c)) as a white powder:2.9 g (79%).

1H NMR (DMSO-D₆, 200MH_(Z)) δ4.49 (2H, d); 7.32 (1H, dd); 7.5-7.7 (3H,m); 8.0-8.2 (2H, m); 8.42-8.53 (1H, m); 9.27 (1H, t, NH); 13.0-13.4 (1H,s, COOH).

MS (ESP+) m/z 324 (M+H)⁺, 159.

Anal. Calcd for C₁₅H₁₁NO₃Cl₂.0.4H₂O C, 54.4; H, 3.59; N, 4.23.

Found C, 54.0; H, 3.2; N, 4.2.

1-(3-Dimethylaminopropyl)-3-ethyl carbodiimide.HCl (655 mg, 3.4 mmol)and 1-Hydroxybenztriazole (463 mg, 3.4 mmol) were added portionwise to astirred solution of (18(c)) (1.0 g, 3.1 mmol) in DMF (20 mL) at 0° underargon. After 30 mins a solution of compound (15(b)) (Example 15) (1.13g, 3.57 mmol) in DMF (20 mL) was added dropwise, followed by N-methylmorpholine (375 ml, 3.4 mmol). The mixture was then allowed to warm toroom temperature over 4 hours. The resulting reaction mixture wasconcentrated to 1/5 volume and diluted with EtOAc(100 mL). This solutionwas then washed successively with 1N citric acid (100 mL), saturatedNaHCO₃(aq) (100 mL), water (100 mL) and brine (100 mL), dried overMgSO₄, filtered and concentrated to a white foam. This was then purifiedby flash chromatography on 9385 SiO₂, eluting on a gradient of 50-75%EtOAc/i-Hexane to yield(2S,4S),4-BOCsulfanyl-2-{[3-(3,4-dichlorobenzylcarbamoyl)-benzoylamino]-methyl}-pyrrolidine-1-carboxylicacid allyl ester (18(d)) as a white foam: 1.57 g (82%).

¹H NMR (CDCl₃, 250 MH_(Z)) δ1.5 (9H, s); 1.6-1.9 (1H, m); 2.55-2.75 (1H,m); 3.2-3.6 (2H,m); 3.65-3.9 (2H, m); 4.1-4.25 (2H, m); 4.5-4.65 (4H,m); 5.15-5.35 (2H, m); 5.38-6.0 (1H, m); 6.87 (1H, t, NH); 7.2 (1H, dd);7.4 (1H, d); 7.45 (1H, d); 7.55 (1H, t); 7.95 (1H, d); 8.07 (1H, d);8.25 (1H, s); 8.35-8.6 (1H, s, NH).

MS (ESP+) m/z 622 (M+H)⁺,566.522.

Anal. Calcd for C₂₉H₃₃N₃Cl₂O₆S: C, 55.9; H, 5.34; N, 6.75.

Found C, 56.1; H, 5.6; N, 6.6.

Compound (18(d)) was deprotected (analogously as for the equivalent stepin Example 15) to give the desired starting material (18(e)) in 67%yield.

¹H NMR (CDCl₃,200 MH_(Z)) δ1.2-1.6(10H, m); 2.25-2.55 (2H, m1H+1NH); 2.9(1H,q); 3.3-3.75 (5H, m); 4.6 (2H, d); 6.9-7.05 (1H, m, NH); 7.05-7.15(1H, m, NH); 7.2 (1H, dd); 7.4 (1H, d); 7.45 (1H, d); 7.52 (1H, t);7.9-8.05 (2H, m); 8.23 (1H, m).

MS (ESP+) m/z 538 (M+H)⁺, 438.

EXAMPLE 19

(See Scheme 19)

(2S,4S),4-sulfanyl-2-[(3-methoxycarbonyl-benzoylamino)-methyl]-pyrrolidin-1-carboxylicacid allyl ester

(2S,4S),4-BOCsulfanyl-2-[(3-methoxycarbonyl-benzoylamino)-methyl]-pyrrolidin-1-carboxylicacid allyl ester (19(a)), (300 mg, 0.63 mmol) was dissolved in TFA (5mL) at room temperature under argon. The reaction mixture wasconcentrated and azeotroped with toluene (3×20 mL) to yield the desiredproduct (19) as a colourless viscous gum: 250 mg (105%).

¹H NMR (CDCl₃, 200 MH_(Z)) δ1.6-1.85 (2H, m, CH+SH); 2.55-2.85 (2H, m);3.1-3.6 (3H, m); 3.92 (3H, bs); 4.0-4.4 (2H, m); 4.65 (2H, d); 5.15-5.4(2H, m); 5.8-6.1 (1H, m); 7.53 (1H, t); 8.0-8.1 (1H, m); 8.1-8.25 (1H,m); 8.3-8.7 (2H, m, Aromatic-H+NH).

MS (FAB) m/z 379 (M+H)⁺, 163.

Starting material (19(a)) was prepared as follows. A suspension ofisophthalic acid monomethyl ester (compound 18(a), Example 18). (2.5 g,13.89 mmol) in CH₂Cl₂ (50 mL) and DMF (10 drops) was treated withoxallyl chloride (135 mL, 15.5 mmol) at 0° under argon. The reactionmixture was allowed to warm to room temperature over 18 h. The resultingsolution was concentrated and azeotroped with toluene to give acrystalline yellow solid. This was then redissolved in CH₂Cl₂ (50 mL)and added dropwise to a stirred solution of(2S,4S)-2-aminomethyl-4-BOCsulfanyl-pyrollidine-1-carboxylic acid allylester (compound 15(b), Example 15) (2.0 g, 6.33 mmol) and (^(i)Pr)₂NEt(2.2 mL, 12.66 mmol) in CH₂Cl₂ (50 mL) at 0° under argon. The reactionmixture was allowed to warm to room temperature and stirred for 18hours, then washed with water (2×50 mL), dried over MgSO₄, filtered andconcentrated to a dark brown oil. This was then purified by flashchromatography on 9385 SiO₂ eluting with a gradient of 25-50%EtOAc/i-Hexane to yield the desired starting material (19(a)) as a paleyellow, viscous oil: 1.81 g (60%).

¹H NMR (CDCl₃,200 MH^(Z)) δ1.5 (9H, s); 1.65-1.9 (1H, m); 2.55-2.8 (1H,m); 3.3 (1H, q); 3.4-3.65 (1H, m); 3.65-3.9 (2H, m); 3.95 (3H, s);4.05-4.35 (2H, m); 4.6-4.7 (2H, m); 5.15-5.4 (2H, m); 5.8-6.1 (1H, m);7.52 (1H, t); 8.02 (1H, dd); 8.15 (1H, dd); 8.24-8.5 (1H, bs, NH); 8.55(1H, bs). MS (FAB) m/z 479 (M+H)⁺, 423.163.

Anal. Calcd for C₂₃H₃₀N₂O₇S: C, 57.7; H, 6.32; N, 5.85.

Found C, 57.5; H, 6.4; N, 5.7.

EXAMPLE 20

(See Scheme 20)

N-([2S,4S],4-sulfanyl-pyrrolidin-2yl-methyl)-3-phenoxy-benzamide

3-Phenoxybenzoic acid was coupled with(2S,4S)-2-aminomethyl-4-BOCsulfanyl-pyrollidine-1-carboxylic acid allylester (compound (15(b)), Example 15). followed by selective deprotectionof the N-allyloxycarbonyl group and removal of the BOC group(analogously to the equivalent steps in Example 15) to give the desiredproduct 20.

NMR CDCl₃ δ1.8 (1H, m), 2.72 (1H, m), 3.01-3.31 (1H, bd), 3.69-3.97 (4H,m), 4.3 (1H, bs), 6.92-7.17 (4.5H, m, aromatics), 7.23-7.45 (5.5H, m,aromatics), 7.56 (1H, m), 7.68 (1H, t), 8.02-8.29 (1H, 2t), 9.02-9.29(1H, 2bs). +ether.

Analysis requires for C₁₈H₂₀N₂O₂S.HI C=47.33; H=4.6; N=6.13; FoundC=47.8; H=4.5; N=6.1.

EXAMPLE 21

(See Scheme 21)

5-{([2S,4S],1-allyloxycarbonyl-4-sulfanyl-pyrrolidin-2yl-methyl)-carbamoyl}-isophthalicacid dimethyl ester

Benzene-1,3,5-tricarboxylic acid dimethyl ester was coupled to(2S,4S)-2-aminomethyl-4-BOCsulfanyl-pyrollidine-1-carboxylic acid allylester (compound (15(b)), Example 15), followed by removal of the BOCgroup (analogously to the equivalent steps in Example 15) to give thedesired product 21.

NMR CDCl₃ δ1.67 (1H, m), 1.75 (1H, d), 2.66-2.89 (3H, m), 3.21 (1H, q),3.27-3.37 (1H, m), 3.5 (1H, m), 3.9 (2H, bs), 3.97 (6H, s), 4.08-4.27(2H, m), 4.68 (2H, d), 5.2-5.4 (2H, m), 5.88-6.06 (1H, m), 8.68 (2H,bs), 8.8 (1H, d).

Analysis requires for C₂₀H₂₄N₂O₇S, C=55.0; H=5.54; N=6.42; Found C=54.9;H=5.6; N=5.75.

EXAMPLE 22

(See Scheme 22)

(2S)-2-{3[([2S,4S]-4-sulfanyl-pyrrolidin-2-yl-methyl)amino]-benzoyl-amino}-4-methylsulfanyl-butyricacid methyl ester

(2S)-2-{3-[([2S,4S]-4-BOCsulfanyl-pyrrolidin-2-yl-methyl)-amino]-benzoylamino}-4-methylsulfanyl-butyricacid methyl ester (22 g) was deprotected (analogously as for theequivalent step in Example 15) to yield the desired end product (22).

¹H NMR (CDCl₃+CD₃COOD) δ1.7-1.9(1H,m);2.0-2.4(6H+CH₃COOH,M);2.5-2.8(3h,M);3.23(1h,Q);3.45-3.7(2H,m);3.7-3.9(4H,m);3.95-4.15(1H,m);4.8-4.95(1H,m);6.8(1H,d);7.05-7.18(2H,m);7.23(1H,t).

MS (ESP) m/z 398 (M+H)⁺,235.

Anal.Calcd for C₁₈H₂₇N₃O₃S₂1.25TFA: C,45.6;H,5.27;N,7.78.

Found C,45.2;H,5.3;N,7.4.

Starting material 22 g was prepared as follows.

i) Preparation of(2S,4S),4-BOCsulfanyl-2-formyl-pyrrolidine-1-carboxylic acid allyl ester(22b)

TPAP (5.5 mg, 0.0156 mmol) was added to a stirred mixture of(2S,4S),4-BOCsulfanyl-2-hydroxymethyl-pyrrolidine-1-carboxylic acidallyl ester (22a)(100 mg, 0.31 mmol) and NMM-O (56 mg, 0.478 mmol) inCH₂Cl₂(2.0 mL) and CH₃CN (100 μL) containing dried powdered 4A°molecular sieve(200 mg). The reaction mixture was left to stir for 1 hthen concentrated to dryness. This was then purified by flashchromatography on SiO₂ (Varian Mega Bond Elut Column) eluting with 50%EtOAc/i-Hexane to give compound 22b as a colourless gum: 66.3 mg(66.7%).

¹H NMR (CDCl₃,250 MH_(Z)) δ1.4-1.6(9H,m);2.0-2.25(1H,m);2.45-2.75(1H,m);3.45-3.6(1H,m);3.75-3.9(1H,m);3.9-4.1(1H,m);4.1-4.35(1H,m);4.5-4.7(2H,m);5.15-5.4(2H, m);5.75-6.05(1H,m);9.4(1H,s,CHO).

MS (CI) m/z 316 (M+H)⁺,260.216.

ii) Preparation of(2S),2-[(3-amino-benzoyl)-amino]-4-methylsulfanyl-butyric acid methylester (22e)

3-Nitro-benzoic acid (22c)(2.0 g, 11.9 mmol) was coupled withL-methionine methyl ester hydrochloride (2.6 g, 13 mmol) according tothe method used to synthesise compound 18a, to give(2S),2-[(3-nitro-benzoyl)-amino]-4-methylsulfanyl-butyric acid methylester (22d) as a white solid: 3.15 g(93.4%)

¹H NMR (CDCl₃,200 MH_(Z))Δ2.05-2.45(5H,m);2.63(2H,t);3.82(3H,s);4.96(1H,m); 7.2(1H,d,NH);7.65,1H,t);8.18(1H,m);8.39(1H,m);8.65(1H,m).

MS (ESP) m/z 313 (M+H)⁺, 265.253.

Anal. Calcd for C₁₃H₁₆N₂O₅S: C,50.0;H,5.516;N,8.97.

Found C,50.3;H,5.1;N,8.9.

A stirred solution of 22d (500 mg, 1.62 mmol) in MeOH(10 mL) was treatedportionwise with decolourising charcoal (50 mg), and iron III chloridehexahydrate (7 mg, 0.026 mmol). N,N-Dimethyl hydrazine (1.5 mL, 19.8mmol) was then added dropwise and the resulting suspension was heated toreflux for a total of 18 h. The reaction mixture was then concentratedto dryness and the residues purified by flash chromatography on SiO₂(Varian Mega Bond Elut Column) eluting with 50% EtOAc/i-Hexane. Productfractions were then concentrated to yield a colourless oil whichcrystallised on standing. This was then triturated with Et₂O to give 22eas a white powder which was collected by filtration and dried: 367 mg(81.2%)

¹H NMR (CDCl₃,250 MH_(Z)) δ2.0-2.4(5H,m);2.5-2.65(2H,m);3.8(3H,s);4.9(1H,m); 6.75-6.95(2H,m,ArH+CONH);7.05-7.3(3H,m).

MS (ESP) m/z 283 (M+H)⁺, 251,235,23.

Anal. Calcd for C₁₃H₁₈N₂O₃S: C,55.3;H,6.43;N,9.92.

Found C,55.5;H,6.6;N,9.8.

iii) Preparation of 22g

A solution containing 22e (50 mg, 0.17 mmol) and 22b (54 mg, 0.17 mmol)in EtOH(2.5 mL) was treated with powdered 4A° molecular sieves (100 mg)and the resulting suspension was stirred at room temperature for 1 h.Acetic acid (10 μL) and sodium cyanoborohydride(17 mg, 0.27 mmol) werethen added and the reaction mixture was left to stir for 18 h at roomtemperature. The reaction mixture was then partitioned between EtOAc(50mL) and saturated NaHCO₃(aq)(50 mL). The aqueous phase was then washedwith EtOAc(50 mL) and the combined organics dried over MgSO₄, filteredand concentrated to a colourless gum. This was then purified by flashchromatography on SiO₂ (Varian Mega Bond Elut Column) eluting a gradientof 25-40% EtOAc/i-Hexane to give(2S)-2-{3-[([2S,4S]-1-allyloxycarbonyl-4-BOCsulfanyl-pyrrolidin-2-yl-methyl)-amino]-benzoyl-amino}-4-methylsulfanyl-butyricacid methyl ester (22f) as a colourless gum: 60.1 mg(60.3%).

1H NMR (CDCl₃,200 MH_(Z)) δ1.45(9H,s,^(t)Bu); 1.7-1.9(1H,m);2.0-2.4(5H,m); 2.45-2.7(3H,m); 3.1-3.35(2H,m); 3.4-3.6(1H,m);3.6-3.85(4H,m); 4.0-4.3(2H,m); 4.6(2H,m); 4.8-4.95 (1H,m);5.15-5.4(2H,m); 5.8-6.1(1H,m); 6.75(1H,d); 6.5-7.3(5H,m).

MS (ESP) m/z 582 (M+H)⁺,482.

Compound 22f was deprotected (analogously as for the equivalent step inExample 15) to give the desired starting material 22 g in 64% yield.

¹H NMR (CDCl₃+D₂O) δ1.15-1.95 (10H,m); 1.95-2.15(4H,m,SMe+H);2.15-2.35(1H,m); 2.35-2.5(1H,m); 2.55(2H,t); 2.75-2.95(1H,m);2.95-3.15(1H,m); 3.15-3.55(3H,m); 3.55-3.7(1H,m); 3.78(3H,s,COMe);4.9(1H,m); 6.73(1H,m); 6.98-7.13(2H,m);7.2(1H,t).

MS (ESP) m/z 498 (M+H)⁺,398.

Anal.Calcd for C₂₃H₃₅N₃O₅S₂O.35CH₂Cl₂: C,53.2;H,6.82N,7.97.

Found C,53.5;H,7.1;N,7.5.

EXAMPLE 23

(See Scheme 30)

Preparation ofN-((2S,4S)-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-N-(2-naphthalen-1-yl-ethyl)butyramide(compound 9);(2S,4S)-2-{[(3-Methoxypropyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol(compound 10) and;(2S,4S)-2-{[(2-(4-Methoxyphenyl)methyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol(compound 11).

Preparation of Compound 9

A solution of starting materialN-((2S,4S)-4-BOCsulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-N-(2-naphthalen-1-yl-ethyl)butyramide(6) (770 mg) in trifluoroacetic acid (40 ml) was stirred at ambienttemperature for 10 minutes. The trifluoroacetic acid was evaporatedunder reduced pressure and the residue redissolved in diethyl ether (90ml). Ethereal HCl(1M, 10 ml) was added and the resulting suspensioncentrifuged. The diethyl ether was decanted off and more ether(90 ml)added to the residue. This mixture was stirred for five minutes and thenrecentrifuged. The washing/centrifuging procedure was repeated once moreand the resulting white solid dried under reduced pressure to givecompound (9), (600 mg)

NMR, data in DMSOd6 d 0.6(2d, 6H), 0.95(d, 1H), 1.7(m, 3H), 2.15(m, 1H),1.9(m, 1H), 3.0 to 3.85(m, 10h), 7.3 to 8.4(m, 7H), 8.9(br,s, 1H),9.5(br,s, 1H).

Micro Analysis: %Theory C,64.9; H,7.7; N,6.9. (1.00 HCl) %Found C,64.7;H,7.9; N,6.8.

Starting material (6) was prepared as follows.

(2S,4S)-2-Formyl-4-BOCsulfanyl-pyrrolidine-1-carboxylic acid allyl ester(1) (1.84 g) in dichloromethane(20 ml) was added dropwise over 10minutes to a mixture of 2-naphthalen-1-ylethylamine (1.0 g),. sodiumtriacetoxyborohydride(1.36 g) and 4A powdered molecular sieve (3.0 g) indichloromethane (130 ml) cooled to −20° C. and stirred under an argonatmosphere. After the addition was complete the reaction was allowed towarm to ambient temperature and stirred for a further 18 hours. Themolecular sieves were filtered off and the fitrate stirred withsaturated aqueous sodium bicarbonate solution( 100 ml) for 5 minutes.The mixture was separated, the organic phase dried over magnesiumsulphate and applied to a silica flash column which was then eluted with1.Ethyl acetate/Hexane(50:50), 2.Ethyl acetate/Hexane(80/20), 3.Ethylacetate to give(2S,4S)-4-BOCsulfanyl-2[(2-naphthalen-1-ylethylamino)-methyl]pyrrolidine-1-carboxylicacid allyl ester (2) (2.2 g) as a colourless gum.

NMR data in CDCl₃, d 1.5(s, 9H), 1.85(m, 1H), 2.5(m, 1H), 2.8(m, 1H),3.0(m, 3H), 3.2(m, 3h), 3.7(m, 1H), 4.05(m, 2H), 4.55(d, 2H), 5.25(m,2H), 5.9(m, 1H), 7.43(m, 4H), 7.7(d, 1H), 7.83(m, 1H), 8.05(m, 1H).

A mixture of compound (2)(1.2 g), isovaleryl chloride(0.61 g) andtriethylamine(0.77 g) in dichloromethane(75 ml) was stirred for 1 hourat ambient temperature. The reaction mixture was then applied to asilica flash colomn which was eluted with ethyl acetate/hexane(20:80) togive compound(3) as a colourless gum (1.3 g).

Tributyltin hydride(6.46 g) was added dropwise over 5 minutes to astirred mixture of compound(3)(1.23 g) andbis(triphenylphosphine)palladium(0) chloride(20 mg) indichloromethane(75 ml). This mixture was stirred at ambient temperaturefor 30 minutes and then applied to a silica flash column which waseluted with 1.Ethyl acetate/Hexane(50:50), 2.Ethyl acetate, 3.Ethylacetate/Methanol(95:5). The product obtained was recolumned on anIsolute® C18(10 g) column eluting with methanol/water(80:20) to givestarting material compound (6) as a white solid (769 mg), m.pt. 86°.

NMR data (CDCl₃) d 0.9(2d, 6H), 1.3(m, 1H), 1.5(s, 9H), 1.8-2.5(m, 6H),2.9(m, 1H), 3.05-3.9(m, 9H), 7.25-8.35(m, 7H).

Preparation of Compound(10)

A solution of starting material(2S,4S-2-{[(3-methoxypropyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4BOCthiol(compound 7) (78 mg) in trifluoroacetic acid(5 ml) was stirred atambient temperature for 30 minutes. The trifluoroacetic acid was removedunder reduced pressure and the residue treated with diethyl ether(5 ml).The ether was decanted off and the residue dried under reduced pressurefor 24 hours to give the desired end product as a colourless gum(compound 10)(70 mg).

NMR data (CDCl₃) d 1.95(m, 4H), 2.05(m, 1H), 3.16-3.62(m, 10H), 3.29(s,3H), 3.7(m, 1H), 4.15(m, 2H), 7.3-7.65(m, 4H), 7.68((d, 1H), 7.88(d,1H), 7.98(d, 1H), 11.2(br,s, 2H).

Micro Analysis: %Theory C,48.2; H,5.13; N,4.32. (2.5TFA, 0.25H₂O) %FoundC,48.5; H,5.20; N,4.40.

Starting material (compound 7) was prepared as follows.

A solution of 4-methoxy-butyraldehyde(140 mg) in dichloromethane(10 ml)was added dropwise to a mixture of compound (2)(250 mg), sodiumtriacetoxyborohydride(338 mg) and 4A molecular sieves(1.0 g) indichloromethane(30 ml) stirred under an argon atmosphere at −20°. Afterthe addition was completed (5 minutes) the reaction mixture was allowedto warm to ambient temperature and stirred for 18 hours. The molecularsieves were filtered off and the filtrate washed with saturated sodiumbicarbonate solution(20 ml), then brine and dried over magnesiumsulphate. The solution was then applied to a silica column and elutedwith ethyl acetate/hexane(50:50) to give a clear gum, compound(4)(260mg).

Compound(7) was synthesised from compound(4) analogously to thepreparation of compound(6).

NMR data (CDCl₃) d 1.35(m, 1H), 1.48(s, 9H), 1.74(m, 2H), 2.31(m, 1H),2.42-3.1(m, 7H), 3.15-3.5(m, 9H), 3.65(m, 1H), 7.28-8.1(m, 7H).

Preparation of Compound(11)

Compound(11) was synthesised from starting material(2S,4S)-2-{[(2-(4-methoxyphenyl)methyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-BOCthiol(compound 8) by the method described for the equivalent step inpreparation of compound(10).

NMR data (CDCl₃) d 1.9(m, 1H), 2.05(m, 1H), 2.3(m, 1H), 3.1-3.8(m, 8H),3.82(s, 3H), 4.25(m, 3H), 6.96(d, 2H), 7.42(m, 6H), 7.83(m, 3H).

Micro Analysis: %Theory C,55.7; H,5.77; N,4.06. (2TFA, 0.75diethylether) %Found C,56.0; H,5.40; N,4.50.

The starting material for compound(11) was prepared as follows;

A mixture of compound(2) (200 mg), p-methoxybenzyl chloride(133 mg),saturated aqueous sodium bicarbonate(5 ml) and dichloromethane(20 ml)was stirred at ambient temperature for 24 hours. The layers wereseparated and the organic layer dried, applied to a silica flash columnwhich was then eluted with ethyl acetate/hexane(80:20) to give(2S,4S)-1-allyloxycarbonyl-2-{[(2-(4-methoxyphenyl)methyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-BOCthiolcompound(5) as a colourless gum(140 mg).

NMR data (CDCl₃) d 1.45(s, 9H), 2.0(m, 1H), 2.35(m, 1H), 2.53-4.15(m,10H), 3.8(s, 3H), 4.6(m, 4H), 5.25(m, 2H), 5.9(m, 1H), 6.85(m,3H),7.3(m, 6H), 7.75(m, 2H).

The desired starting material (compound(8)) was synthesised fromcompound(5) by the same procedure used to prepare compound(6) fromcompound (3).

Mass Spec.(ESP+) m/e 507.0

EXAMPLE 24

(See Scheme 31)

Preparation of a)3-Methyl-N-(naphthalen-1-ylmethyl)-N-([2S,4S]4-sulfanylpyrrolidin-2-ylmethyl)-butanamide(compound 23); b)N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-pentanamide(compound 24); c)N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-2-(pyridin-3-yl)-acetamide(compound 27); d)3-Methyl-N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-pentanamide(compound 25); e)3-Methoxy-N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-propanamide(compound 26) and; f)(2S,4S)-2-[{N-(4-methoxybenzyl)-N-(naphthalen-1-ylmethyl)-amino}-methyl]-pyrrolidine-4-thiol(compound 54). a) Preparation of Compound 23

A solution of starting material3-methyl-N-(naphthalen-1-ylmethyl)-N-([2S,4S]4-BOCsulfanyl-pyrrolidin-2-ylmethyl)-butanamide(compound(18)) (187 mg) in trifluoroacetic acid (10 ml) was stirred atambient temperature for 5 minutes. The trifluoroacetic acid wasevaporated under reduced pressure and the resulting residue wasredissolved in ethyl acetate (5 ml). A solution of hydrogen chloride (2ml/1.0M) was added to the solution followed by diethylether (5 ml). Themixture was centrifuged, the solvent decanted off and the residue waswashed with more diethylether (2×15 ml) and dried to give thehydrochloride salt of compound(23) as an off-white solid (43 mg).

N.M.R. data (DMSO-d6) δ0.83 (m,6H), 0.95(d,1H), 1.68(m,1H), 2.10(m,3H),2.42(m,1H), 3.10(m,1H), 3.28-3.90(m,5H), 5.20(m,2H), 7.08(d,1H),7.57(m,3H), 7.87(d,1H), 8.00(m,2H), 9.10-9.80(2br,s,2H)

Micro Analysis: Theory % C,62.7; H,7.52; N,6.97. (1HCl.0.5H₂O) Found %C,62.4; H,7.6; N,6.7.

The starting material compound(18) was prepared as follows.

A solution of (2S,4S)-2-formyl-4-BOCsulfanyl-pyrrolidine-1-carboxylicacid allyl ester (compound (1)) (3.11 grm. ) in dichloromethane(60 ml.)was added dropwise to a stirred mixture of of 1-naphthalenemethylamine(1.71 g), 4A molecular sieves(12 grms) and sodiumtriacetoxyborohydride(2.3 grms) in dichloromethane (200 ml) under anargon atmosphere at −20°. The mixture was stirred for a further 30minutes at −20° C. and then allowed to warm to ambient temperature andstirred for a further 16 hours. The mixture was filtered and washed withaqueous sodium bicarbonate solution (2×200 ml),the organic phase furtherwashed with water (200 ml), separated, dried over magnesium sulphate andpurified by column chromatography, using ethyl acetate/hexane (30:70) aseluent to give(2S,4S)-2-{[naphthalen-1-ylmethyl]-amino)-methyl}-4-BOCsulfanyl-pyrrolidine-1-carboxylicacid allyl ester (compound(12)) as a pale yellow oil (2.09 g).

N.M.R. data (CDCl₃) δ1.50(s,9H), 1.55(m,1H), 1.90(m,1H), 2.50(m,1H),2.90(m,1H), 3.05(m,1H),3.20(m,1H), 3.68(m,1H), 4.08(m,2H), 4.23(s,2H),4.55(d,2H), 5.20(m,2H), 5.90(m,1H), 7.47(m,4H), 7.77(m,1H), 7.86(m,1H),8.13(m,1H).

A mixture of compound(12) (507 mg), triethylamine(0.3 ml) and isovalerylchloride(0.271 ml) in dichlorometane (30 ml) was stirred at ambienttemperature for 1.5 hours and then applied directly to a silica flashcolumn. This was eluted with ethyl acetate/hexane (25:75) andethylacetate/hexane(35:65) to give3-Methyl-N-(naphthalen-1-ylmethyl)-N-([2S,4S]-1-allyloxycarbonyl-4-BOCsulfanylpyrrolidin-2-ylmethyl)-butanamide(compound(13)) as a gum (475 mg).

N.M.R. data (DMSO-d6. 373° K) δ0.90(m,6H), 1.45(s,9H), 1.78(m,1H),2.18(m,3H), 2.50(m,1H), 3.15(q,1H), 3.45(m,1H), 3.70(m,2H), 4.03(q,1H),4.20(m,1H), 4.45(m,2H), 5.10(m,4H), 5.80(m,1H), 7.20(d,1H), 7.50(m,3H),7.80(d,1H), 7.92(m,1H), 8.00(m, 1H).

Tributyltin hydride(2.22 ml) was added dropwise to a mixture ofcompound(13) (446 mg), bis-triphenylphosphine palladium chloride(5.8 mg)in dichloromethane (10 ml). The mixture was stirred at ambienttemperature under an argon atmosphere for 70 minutes and then applieddirectly to a flash column which was eluted with (1)Ethyl acetate/hexane(50:50) and (2) Ethyl acetate. The product obtained was recolumned on anIsolute® C18 (10 g) column, eluting with methanol/water (1) (70:30),(2)(75:25) and (3)(80:20) to give the desired starting material(compound(18)) as a gum (197 mg).

N.M.R. data (DMSO-d6,373° .K) δ0.90(m,6H), 1.45(m,5H), 1.60(m,1H),1.68(m,2H), 2.12(m,2H), 2.25(d,2H), 2.40(m,1H), 2.60-3.85(m,8H),5.14(s,2H), 7.20(d,1H), 7.50(m,3H), 7.83(m,1H), 7.93(m,1H), 8.03(m,1H).

b) Preparation of Compound 24

Compound(24) was synthesised by the same procedure used for compound(23)but substituting appropriate compounds as indicated in Scheme 31.

Compound 24

N.M.R. data (DMSO-d6) δ0.85(m,3H), 1.15-1.75(m,5H), 2.28(t,2H),3.10(m,1H), 3.33-3.95(m,6H), 5.18(m,2H), 7.20(2d,1H), 7.55(m,3H),7.85(d,1H), 8.00(m,2H), 8.94-9.90(2br,s,2H)

Micro Analysis: %Theory C,62.7; H,7.52; N,6.97. (1HCl.0.5H₂O) %FoundC,62.5; H,7.80; N,6.8.

Compound(14):

N.M.R. data (CDCl₃) δ0.90(m,3H), 1.12-2.10(m,6H), 1.48(s,9H),2.26(m,1H), 2.50(m,1H), 3.00-5.70(m,12H), 5.87(m,1H), 7.07-8.06(m,7H).

Compound(19):

N.M.R. data (DMSO-d6.373° .K) δ0.84(m,3H), 1.30(m,3H), 1.45(s,9H),1.55(m,2H), 2.34(m,3H), 2.80(m,2H), 3.45(m,5H), 5.10(m,2H), 7.25(d,1H),7.50(m,3H), 7.90(m,1H), 8.03(m,1H).

c) Preparation of Compound(27)

Compound(27) was synthesised in the same manner as the equivalent stepfor compound(23), from starting materialN-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-BOCsulfanylpyrrolidin-2-ylmethyl)-2-(pyridin-3-yl)-acetamide(compound(22)).

Compound(27):

N.M.R. data (DMSO-d6) δ1.70(m,1H), 2.50(m,1H), 3.14(m,1H),3.28-5.10(m,7H), 5.35(m,2H), 7.20-9.00(m,11H), 9.20(br,s, 1 H),10.05-10.50(2br.s.1H)

Micro Analysis: %Theory C,55.10; H,6.60; N,7.97. (2HCl,2.25H₂O, 0.3diethyl ether) %Found C,54.80; H,6.10; N,7.60.

Starting material (compound(22)) was synthesised as follows.

A mixture of compound(12)(345 mg), 4dimethylamino-pyridine(305 mg),3-pyridylacetic acid hydrochloride(262 mg) and1-(3-dimethylaminopropyl-3-ethylcarbodiimide hydrochloride(348 mg) indichloromethane (30 ml) was stirred at ambient temperature, under anargon atmosphere, for 16hours. The mixture was then purified by silicaflash column chromatography, eluting with ethyl acetate/hexane(75:25)and then ethyl acetate to giveN-(naphthalen-1-ylmethyl)-N-([2S,4S]-1-allyloxycarbonyl-4-BOCsulfanylpyrrolidin-2-ylmethyl)-2-pyridin-3-yl)-acetamide(compound(17)) as a colourless gum (394 mg).

Compound(17)

N.M.R. data (DMSO-d6, 373° K) δ1.46(s,9H), 1.75(m,1H), 2.50(m,1H),3.17(q,1H), 3.50(m,1H), 3.75(m,4H), 4.04(m,1H), 4.27(m,1H), 4.45(m,2H),5.15(m,4H), 5.83(m,1H), 7.25(m,2H), 7.43(t,1H), 7.52(m,2H), 7.58(m,1H)7.82(d,1H), 7.95(m,2H), 8.40(d,2H).

Using the procedure previously described for the equivalent step insynthesis of compound 23, the desired starting material (compound(22))was synthesised from compound(l7).

Compound(22)

N.M.R. data(DMSO-d6, 373° K) δ1.45(s,9H), 2.38(m,1H), 2.55-4.00(m,10H),5.20(m,2H), 7.25(m,2H), 7.50(m,4H), 7.90(m,3H), 8.40(m,2H).

d) Preparation of Compound(25)

Compound(25) was synthesised using compounds 12, 15 and 20 asintermediates, in the same manner as the equivalent steps for synthesisof compound (27) (see Scheme 31).

Compound(25)

N.M.R. data (DMSO-d6) δ0.80(m,6H), 0.95-4.80(m,14H), 5.18(m,2H),7.08(d,1H), 7.55(m,3H), 7.95(m,3H), 8.90-10.15(2br,d,2H).

Micro Analysis: %Theory C,59.1; H,7.30; N,6.27. (2HCl, 0.2H₂O) %FoundC,59.1; H,6.90; N,5.9.

Compound(15);

N.M.R. data (DMSO-d6, 373° K) δ0.85(m,6H), 1.15(m,1H), 1.35(m,1H),1.45(s,9H), 1.75(m,1H), 1.90(m,1H), 2.17(m,1H), 2.30(m,1H), 2.50(m,1H),3.15(q,1H), 3.45(m,1H), 3.70(m,2H), 4.03(q,1H), 4.20(m,1H), 4.44(d,2H),5.10(m,4H), 5.80(m,1H), 7.20(d,1H), 7.50(m,3H), 7.80(d,1H), 7.90(m,1H),8.00(m,1H).

Compound(20):

N.M.R. data (DMSO-d6, 373° K) δ0.85(m,6H), 1.25(m,3H), 1.45(s,9H),1.93(m,1H), 2.27(m,3H), 3.40(m,6H), 5.13(m,2H), 7.25(d,1H), 7.50(m,3H),7.80(d,1H), 7.90(m,1H), 8.04(m,1H).

e) Preparation of Compound(26)

Compound(26) was synthesised using compounds 12, 16 and 21 asintermediates in the same manner as the equivalent steps for synthesisof compound(27) (see Scheme 31).

Compound(26);

N.M.R. data (DMSO-d6) δ1.70(m,1H), 2.40-4.15(m,14H), 5.20(m,2H),7.20(2d,1H), 7.55(m,3H), 7.85(m,1H), 8.00(m,2H), 9.05-10.25(2br,d,2H).

Micro Analysis: %Theory C,59.5; H,6.99; N,6.93. (2HCl, 0.2H₂O) %FoundC,59.3; H,7.30; N,6.70.

Compound(16);

N.M.R. data (DMSO-d6, 373° K) δ145(s,9hH), 1.78(m,1H), 2.40-3.80(m,12H),4.00(m,1H), 4.20(m,1H), 4.45(m,2H), 5.10(m,4H), 5.80(m,1H), 7.20(d,1H),7.45(t,1H), 7.50(m,2H), 7.80(d,1H), 7.90(m,1H), 8.00(m,1H).

Compound(21):

N.M.R. data (DMSO-d6, 373° K) δ1.30(m,1H), 1.48(s,9H), 2.30(m,1H),2.56-3.70(m,14H), 5.15(m,2H), 7.30(d,1H), 7.47(t,1H), 7.53(m,2H),7.83(d,1H), 7.94(m,1H), 8.05(m,1H).

f) Preparation of Compound (54)

A mixture of starting material(2S,4S)-2-[{(N-(4-methoxybenzyl)-N-(naphthalen-1-ylmethyl)-amino}-methyl]-pyrrolidine-4-BOCthiol(compound(53))(100 mg) and trifluoroacetic acid(5 ml) was stirred atambient temperature for 1 hour. The trifluoroacetic acid was removedunder reduced pressure and the residue coevaporated with diethylether togive compound(54) as a colourless gum (83 mg).

NMR data (CDCl₃) δ1.5(m, 1H), 1.75(br,d, 1H), 1.95(m, 1H), 2.6(t, 1H),3.05(m, 1H), 3.2(d, 1H), 3.35(m, 2H), 3.85(s, 3H), 4.2(s, 2H), 4.6(2d,2H), 6.95(d, 2H), 7.4(d, 2H), 7.6(m, 4H), 7.9(m, 3H).

Micro Analysis: %Theory C,52.0; H,5.40; N,3.90. (2.5TFA, 0.4 diethylether) %Found C,52.0; H,4.92; N,3.96.

The starting material was prepared as follows.

A mixture of compound(12)(240 mg), dimethylformamide(20 ml), anhydrouspotassium carbonate(80 mg) and p-methoxybenzylchloride(0.143 ml) wasstirred at 70° under an argon atmosphere for 4 hours. The solvent wasremoved under reduced pressure and the residue purified by columnchromatography eluting with ethyl acetate/hexane(20:80) to give acolourless gum(2S,4S)-1-allyloxycarbonyl-2-[{N-(4-methoxybenzyl)-N-(naphthalen-1-ylmethyl)-amino}-methyl]-pyrrolidine-4-BOCthiol(compound(52)) (213 mg).

NMR data (CDCl₃) d 1.45(s, 9H), 2.15(m, 1H), 2.5(m, 1H), 2.8(m, 1H),3.05(m, 1H), 3.5(m, 2H), 3.8(br,s, 7H), 3.9(m, 1H), 4.2(m, 1H), 4.6(s,2H), 5.25(m, 2H), 5.9(m, 1H), 6.85(d, 2H), 7.2(d, 2H), 7.4(m, 4H),7.8(2d, 2H), 8.1(d, 1H).

Tributyltin hydride(0.77 ml) was added to a mixture of compound(52) andbis(triphenyl phosphine) palladium (O) chloride(2 mg) indichloromethane(10 ml). The solution was stirred at ambient temperaturefor 30 minutes. A second portion of tributyltin hydride(0.335 ml) andbis(triphenylphosphine) palladium (O) chloride(2 mg) were added and thestirring was continued for a further 30 minutes. The mixture was applieddirectly to a silica flash column which was eluted with ethylacetate/hexane(25:75),(50:50) and finally ethyl acetate. The productobtained was further purified by reverse phase HPLC on a C18 columneluting with water/methanol/TFA(20:80:0.2) to give the desired startingmaterial (compound(53)) as a colourless gum. (168 mg.)

NMR data (CDCl₃) d 1.45(s, 9H), 1.55(m, 1H), 2.0(m, 1H), 2.5(m, 1H),3.1(d, 1H), 3.4(m, 3H), 3.6(t, 1H), 3.8(s, 3H), 4.1(2d, 2H), 4.4(d, 1H),4.6(d, 1H), 6.95(d, 2H), m 7.4(d,2H), 7.5(m, 4H), 7.9(m, 3H).

Micro Analysis: %Theory C,54.4; H,5.40; N,3.70. (2TFA) %Found C,55.0;H,5.31; N,3.89.

EXAMPLE 25

(See Scheme 32)

Preparation of a)(2S,4S)-2[(N-methylnaphthalen-1-ylamino)methyl]-sulfanylpyrrolidine(compound 36) and: b)N-(naphthalen-1-yl)-N-((2S,4S)-4-sulfanylpyrrolidin-2-yl-methyl)-3-methylbutanamide(compound 37). Preparation of Compound 36

A mixture of starting material(2S,4S)-2[(N-methylnaphthalen-1-ylamino)-methyl]-4-BOCsulfanylpyrrolidine(compound (34)) (110 mg) and trifluoroacetic acid (5 ml) was stirred atambient temperature for 1 hour. The trifluoroacetic acid was removedunder reduced pressure and the residue dried under high vacuum to givecompound(36) as a colourless gum(l 10 mg).

N.M.R. data (CDCl₃) δ1.7 (m,1H), 1.9 (d,1H), 2.6 (m,1H), 2.95 (s,3H),3.1 (2d,1H), 3.5 (m,1H), 3.65 (m,3H), 4.05 (m,1H), 7.0 (br, s,1H), 7.4(t,1H), 7.55 (m,3H), 7.7 (d,1H), 7.85 (m,1H), 8.2 (m,1H).

Micro Analysis: %Found C, 45.5; H, 4.2; N, 5.0. (2.0OTFA. 1.0H₂O)%Theory C, 46.3; H, 4.67; N, 5.4.

The starting material for compound(36) was prepared as follows:

A mixture of (2S,4S)-2-formyl-4-BOCsulfanyl-pyrrolidine-1-carboxylicacid allyl ester (compound(1)) ( 711 mg), ethanol(25 ml),1-naphthylamine(333 mg) and 3A molecular sieves(4.5 g.) was stirredunder an argon atmosphere at ambient temperature for 6 hours. Aceticacid (0.4 ml) was added followed by sodium cyanoborohydride(l70 mg). Themixture was then stirred for a further 20 hours when the sieves wereremoved by filtration. The filtrate was concentrated under reducedpressure and the residue applied to a silica column and eluted withethyl acetate/hexane(20:80) to give(2S,4S)-1-allyloxycarbonyl-2[(naphthalen-1-ylamino)-methyl]-4-BOCsulfanylpyrrolidine(compound(31)) as a clear oil (560 mg).

N.M.R. data (CDCl3) δ1.5 (s,9H), 1.85 (m,1H), 2.7 (m,1H), 3.35 (m,2H),3.5 (m,1H), 3.8 (m, 1H), 4.2 (m,1H), 4.5 (m,1H), 4.65 (d,2H), 5.3(2d,2H), 5.95 (m,1H), 6.55 (m,1H), 7.2 (d,1H), 7.3 (t,1H), 7.4 (m,2H),7.75 (m,1H), 7.9 (m,1H).

A mixture of (compound(31))(218 mg), dimethylformamide(40ml),iodomethane(0.6 ml.) and anhydrous potassium carbonate(150 mg) wasstirred at 80° for 20 hours. The solvent was removed under reducedpressure and the residue taken up in ethyl acetate(30 ml.) and washedwith water(20ml). The organic phase was dried over magnesium sulphate,filtered and concentrated under reduced pressure to give(2S,4S)-1-allyloxycarbonyl-2[(N-methylnaphthalen-1-ylamino)-methyl]-4-BOCsulfanylpyrrolidine(compound(32)) as a yellow gum (183 mg).

N.M.R. data (CDCl₃) δ1.45 (s,9H), 2.0 (m,1H), 2.4 (m,1H), 2.85 (s,3H),3.0 (2d,1H), 3.25 (m,1H), 3.7 (2d,1H), 3.8 (m,1H), 4.1 (m,2H), 4.6(d,2H), 5.3 (9m,2h), 5.95 (m,1H), 7.45 (m,5H), 7.8 (m,1H), 8.25 (m,1H).

To a solution of compound(32)(178 mg) in dichloromethane(10 ml) wasadded tri-n-butyltin hydride(0.2 ml.) followed by bis(triphenylphosphine) palladium chloride (2 mg) and the mixture then stirred atambient temperature. After 10 min and 20 min a second and third portionof tri-n-butyltin hydride (0.2ml.) and bis(triphenyl phosphine)palladium chloride (2 mg) were added and stirring continued for afurther 90 min. The reaction solution was applied direct to a silicacolumn and eluted with ethyl acetate/hexane(25:75). (50:50) and ethylacetate. The product was further purified on a reverse phase HPLC. C18column which was eluted with water/methanol/trifluoroaceticacid(20:80:0.2) to give as a colourless gum the desired startingmaterial (compound(34))(160 mg).

N.M.R data (CDCl₃) δ1.45 (s,9H), 2.2 (s,1H), 2.39 (m,1H), 2.85 (s,3H),2.9 (2d,1H), 3.1 (2d,1H), 3.25 (m,2H), 3.4 (m,1H), 3.6 (m,1H), 7.15(d,1H), 7.45 (m,4H), 7.8 (m,1H), 8.35 (m,1H).

Micro Analysis: %Found C, 50.8; H, 5.20; N, 4.6. (2.0OTFA, 0.5H₂O)%Theory C, 49.3; H, 5.13; N, 4.6.

b) Preparation of Compound (37)

A mixture of starting material (compound(35))(187 mg) andtrifluoroacetic acid(5 ml.) was stirred at ambient temperature for 1hour. The trifluoroacetic acid was removed under reduced pressure andthe residue dried under high vacuum to give a colourless gum, compound(37)(200 mg.).

N.M-R. data (CDCl₃) δ0.8 (m,6H), 1.6-2.2 (m,5H), 2.6 (m,1H), 3.2-5.0(m,6H), 7.6 (m,5H), 8.0 (m,2H).

Micro Analysis: %Found C, 48.4; H, 4.80; N, 4.5. (2.0 TFA, 1.0H₂O)%Theory C, 49.0; H, 5.14; N, 4.76.

The starting material was prepared as follows.

Isovaleryl chloride(0.164 ml.) was added dropwise over 10 minutes to astirred solution of compound(31)(297 mg.), dichloromethane(50 ml) andtriethylamine(0.136 ml.). The solution was stirred at ambienttemperature for 24 hours. The solvent was removed under reduced pressureand the residue applied directly to a silica column and eluted withethyl acetate/hexane(25/75) to give a white foam,N-(naphthalen-1-yl-N-((2S,4S)-1-allyloxycarbonyl-4-BOCsulfanylpyrrolidin-2-yl-methyl)-3-methylbutanamide,(compound(33))(329 mg).

N.M.R. data (CDCl₃) δ0.75 (m,6H), 1.5 (s,9H), 1.65 -2.7 (m,5H), 3.15-6.0(m,9H), 7.25 (m,1H), 7.5 (m,3H), 7.7 (m,1H), 7.9 (m,2H).

To a solution of compound(33)(296 mg.) in dichloromethane(10 ml) wasadded tri-n-butyl tin hydride (0.3 ml.) followed by bis(triphenylphosphine) palladium chloride(2 mg.). The solution was stirred atambient temperature. After 10 min and 20 min a second and third portionof tri-n-butyl tin hydride(0.3 ml.) and bis(triphenyl phosphine)palladium chloride(2 mg) were added and the stirring continued for afurther 30 minutes. The reaction solution was applied directly to asilica column which was then eluted with ethyl acetate/hexane(25:75),(50:50) and ethyl acetate. The product was further purified on a reversephase HPLC. C18 column eluting with water/methanol/trifluoroaceticacid(20:80:0.2) to give the desired starting material. (compound(35))(216 mg.).

N.M.R data (CDCl₃) δ0.8 (m,6H), 1.49 (s,9H), 1.1-2.2 (m,6H), 2.9-5.6(m,6H), 7.4-8.0 (m,7H).

Micro Analysis: %Found C, 57.0; H, 6.20; N, 4.80. (1.0TFA, 0.75H₂O)%Theory C, 56.9; H, 6.45; N, 4.91.

EXAMPLE 26

(See Scheme 33)

Preparation of a)3-Methyl-N-(3,3-diphenylpropyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-butanamide(compound 43) and; b)N-(3,3-diphenylpropyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-butanamide(compound 44):

Compounds (43) and (44) were synthesised using the procedure describedin Example 23 using appropriate starting materials and intermediates asset out in Scheme 33.

a) Preparation of Compound (43)

Compound (43)

NMR data (DMSOd6 at 373° K.) d 0.9(d, 6H), 1.7(m, 1H), 2.1(m, IH),2.33(m, 2H), 2.45(m, 1H), 2.9-4.00(m, 9H), 4.2-4.95(m, 2H), 7.3-8.1(m,10H), 9.65(v.br.s, 2H)

Micro Analysis: %Theory C,64.8; H,7.7; N,5.9. 1.00HCl, 1H₂O %FoundC,64.5; H,7.9; N,6.0.

The starting material3-Methyl-N-(3,3-diphenylpropyl)-N-([2S,4S]-4-BOCsulfanylpyrrolidin-2-ylmethyl)-butanamide(compound 41) was synthesised from compound (1) and3,3-diphenylpropylamine using a similar procedure to that outlined inExample 23.

Compound (38):

NMR data (CDCl₃) d 1.5(s, 9H), 1.8(m, 1H), 2.19(m, 2H), 2.42(m, 1H),2.55(m, 2H), 2.7(m, 1H), 2.82(m, 1H), 3.19(m, 1H), 3.67(m, 1H), 4.0(m,3H), 4.55(d, 2H), 5.2(2d, 2H), 5.9(m, 1H), 7.2(m, 10H).

Compound (39):

NMR data (CDCl₃) d 0.75-1.0(m, 6H), 1.22(m, 1H), 1.5(s, 9H),1.78-2.02(m, 2H), 2.3(m, 4H), 3.2(m, 3H), 3.4-4.2(m, 6H), 4.52(m, 2H),5.21(m, 2H), 5.9(m, 1H), 7.2(m, 10H).

Compound (41):

NMR data (CDCl₃) d 0.75-1.00(m, 6H), 1.25(m, 1H), 1.5(s, 9H),1.85-2.4(m, 6H), 2.83(m, 1H), 3.05-3.47(m, 6H), 3.6(m, 1H), 3.87(2t,1H), 7.25(m, 10H).

b) Preparation of Compound (44)

Characterisation data is set out below:

Compound (44):

NMR data (DMSOd6 at 373° K) d 1.65(m, 1H), 1.85(s, 3H), 2.32(q, 2H),2.45(m, 1H), 2.69-4.3(m, 9H), 7.2(m, 10H), 9.37(v.br.s, 2H).

Micro Analysis: %Theory C, 63.3; H, 7.3; N, 6.6. 1.00HCl, 0.75H₂O %FoundC,63.1; H, 7.3; N, 6.7.

Compound (40):

NMR data (CDCl₃) d 1.5(s, 9H), 1.82(s, 3H), 1.6-2.5(m, 4H), 3.2(m, 3H),3.32-4.25(m, 6H), 4.54(m, 2H), 5.23(m, 2H), 5.9(m, 1H), 7.23(m, 10H).

Compound (42):

NMR data (CDCl₃) d 1.48(s, 9H), 1.8(m, 1H), 1.87(s, 2H), 2.07(s, 1H),2.33(m, 3H), 2.83(m, 1H), 3.28(m, 6H), 3.6(m, 1H), 3.85(m, 1H), 7.25(m,10H).

EXAMPLE 27

(See Scheme 34)

Preparation of a)3-Methyl-N-(naphthalen-2-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-butanamide(compound 50) and; b)N-(naphthalen-2-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-acetamide(compound 51)

Compounds (50) and (51) were synthesised using the procedure describedin Example 23 using appropriate starting materials and intermediates asset out in Scheme 34.

a) Preparation of Compound (50)

Compound 50:

NMR data (DMSOd6) d 0.75-1.1(m, 6H), 1.63(m, 1H), 2.1(m, 1H), 2.48(m,1H), 2.83(m, 3H), 3.0-4.95(m, 8H), 7.17(m, 7H).

Micro Analysis: %Theory C,64.2; H,7.44; N,7.13. (1.0HCl) %Found C,64.0;H,7.40; N,7.10.

Starting material3-Methyl-N-(naphthalen-2-ylmethyl)-N-([2S,4S]-4-BOCsulfanylpyrrolidin-2-ylmethyl)-butanamide(compound (48)) was synthesised from compound (1) and2-naphthylmethylamine.

Compound (45):

NMR data (CDCl₃) d 1.48(s, 9H), 1.92(m, 1H), 2.5(m, 1H), 2.82(m, 1H),2.96(m, IH), 3.2(2d, 1H), 3.7(m, 1H), 3.96(s, 2H), 4.08(m, 2H), 4.54(m,2H), 5.2(m, 2H), 5.9(m, 1H), 7.42(m, 3h), 7.8(m, 4H).

Compound (46):

NMR data (CDCl₃) d 0.96(2d, 6H), 1.48(s, 9H), 1.9(m, 1H), 2.13-2.6(m,4H), 3.3(m, 1H), 3.72(m, 2H), 4.15(m, 2H), 4.5(m, 2H), 4.76(m, 1H),5.2(m, 2H), 5.9(m, 1H), 7.48(m, 3H), 7.73(m, 4H).

Compound (48):

NMR data (CDCl₃) d 0.98(2d, 6H), 1.3(m, 1H), 1.48(s, 9H), 2.3(m, 4H),2.9(m, 1H), 3.1-3.7(m, 5H), 4.85(m, 2H), 7.15-7.9(m, 7H).

b) Preparation of Compound (51)

Characterisation data is set out below.

Compound 51:

NMR data (DMSOd6 at 373° K) d 1.7(m, 1H), 2.14(s, 3H), 2.47(m, 1H),2.8-4.00(m, 6H), 4.8(m, 2H), 7.32-8.1(m, 7H).

Micro Analysis: %Theory C,64.2; H,7.44; N,7.13. (1.00 HCl) %FoundC,64.0; H,7.40; N,7.10.

Compound (47):

NMR data (CDCl₃) d 1.5(s, 9H), 1.9(m, 1H), 2.12(s, 2H), 2.29(s, 1H),2.5(m, 1H), 3.18-5(m, 10H), 5.2(m, 2H), 5.95(m, 1H), 7.2-7.89(m, 7H).

Compound (49):

NMR data (CDCl₃) d 1.3(m, 1H), 1.47(s, 9H), 2.15(s, 2H), 2.3(s, 1H),2.35(m, 1H), 2.88(m, 1H), 3.1-3.7(m, 5H), 4.85(m, 2H), 7.4-7.9(m, 7H).

EXAMPLE 28

(See Scheme 35)

(2S)-2-({4-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-2-carbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester (compound 30)

Starting material(2S)-2-({4-[([2S,4S]-4-BOCsulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-2-carbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester 30e (72.1 mg, 0.132 mmol) was deprotected (analogouslyas for the equivalent step in Example 15) to give the title compound 30,76 mg (97.8%).

¹H NMR (CDCl₃+CD₃COOD,200 MHz) d1.75-2.0(1H,m); 2.0-2.5(5H+DMSO,m);2.55-3.0(3H,m); 3.15-3.4(1H,m); 3.5-3.7(1H,m); 3.7-3.9(6H,m);4.2-4.4(1H,m); 4.9-5.05(1H,m); 7.0-8.1(6H,m,ArH).

MS (ESP⁺) m/z 448 (M+H)⁺.

Anal.Calcd for C₂₂ H₂₉ N₃ S₂ O₃ 1.25 TFA C,49.9;H,5.17;N,7.12.

Found C,49.6;H,5.3;N,6.7.

Starting material 30e was prepared as follows.

Compound 30a

2-Napthoic acid was nitrated with conc HNO₃ (Tetrahedron49,17,3655,1993) to give a mixture of nitro-acids 30a, containing therequired 4-Nitro-2-Napthoic acid.

MS (ESP⁻) m/z 216 (M−H)⁻.

Compound 30b

Oxalyl chloride (6.0 mL, 68.7 mmol) was added dropwise to a stirredsolution of the nitro acid mixture,30a(7.3 g, 33.6 mmol) in a mixture ofDMF(1.0 mL) and CH₂Cl₂ (100 mL) at 0° C. under argon. The solution wasallowed to warm to RT stirred 18 hrs, evaporated to dryness andazeotroped with toluene(2×25 mL). The resulting residues wereredissolved in CH₂Cl₂ (100 mL) and cooled to 0° C. under argon. Et₃N(7.0 mL, 50 mmol) was then added, followed by L-Methionine methylester,hydrochloride (7.4 g, 37 mmol), portionwise, such that the internaltemperature did not rise above 10° C. The reaction mixture was left towarm to room temperature and stirred for 18 hr washed with water (100mL), dried over MgSO₄, filtered and concentrated to a viscous brown gum.This was then purified by flash chromatography on SiO₂ (Merck 9385),eluting with 25% EtOAc/i-Hexane. Appropriate fractions were combined andevaporated to give 30b as a viscous orange gum, 490 mg (4%).

¹H NMR (CDCl₃, 200 MHz) d2.1-2.5(5H,m); 2.55-2.75(2H,m); 3.85(3H,s);4.9-5.1(1H,m);7.32(1H,d); 7.6-8.0(2H,m); 8.05(1H,dd); 8.5-8.7(3H,m),

MS (ESP⁺) m/z 363 (M+H)⁺.

Compound 30c

30b (450 mg, 1.24 mmol) was reduced (analogously as for the equivalentstep in Example 22) to give the corresponding aniline, 30c as a yellowgum. 310 mg (75.3%)

¹H NMR (CDCl₃,250 MHz) d2.0-2.45(5H,m); 2.5-2.75(2H,m); 3.83(3H,s);4.3(2H,bs,NH₂); 4.9-5.05(1H,m); 7.0(1H,d,NHCO);7.2(1H,d);7.4-7.65(2H,m); 7.72(1H,s); 7.8-8.0(2H,m).

MS (ESP⁺) m/z 333 (M+H)⁺, 271, 170.

Compound 30d

30c (300 mg, 0.9 mmol) was coupled with the aldehyde 22b(428 mg, 1.36mmol) under the conditions employed to synthesise 22g using MeOH assolvent and in the presence of 3A° molecular sieves as drying agent togive 30d as yellow gum. 460 mg (76.5%)

MS (ESP⁺) m/z 632 (M+H)⁺.

Compound 30e

30d (450 mg, 0.7 mmol) was deprotected (analogously as for theequivalent step in Example 15) to give the desired starting material30e. 220 mg (56.4%)

¹H NMR (CDCl₃,200 MHz) d1.4-1.9(10H+H₂O,m); 2.0-2.75(9H,m); 2.95(1H,q);3.1-3.35(1H,m); 3.35-3.55(2H,m); 3.55-3.8(2H,m); 3.82(3H,s,CO₂Me);4.98(1H,m); 5.15(1H,bs,NH); 6.9-7.1(2H,m,ArH+NHCO); 7.4-7.6(2H,m);7.61(1H,d); 7.8-8.0(2H,m).

MS (ESP⁺) m/z 548 (M+H)⁺, 448.

EXAMPLE 29

(See Scheme 36)

Preparation of a)(2S)-2-({3-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester (compound 31) b)(2S)-2-({3-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl}-amino)-4-methylsulfanylbutyricacid (compound 31f) a) Preparation of Compound 31

31e (55 mg, 0.1 mmol) was deprotected (analogously as for the equivalentstep in Example 15) then treated with Et₂O.HCl to give the titlecompound.31 as a white solid. (37 mg, 64.8%)

¹H NMR (DMSO-D₆+CD₃CO₂D,250 MHz) d1.05(1H,t, (CH ₃CH₂)₂O);1.6-1.8(1H,m); 1.9-2.15(4H,m);2.3-2.7(4H+DMSO,m);3.0-4.0(9H+(CH₃CH₂)₂O); 4.55-4.7(1H,m);6.95(1H,s);7.1(1H,s);7.15(1H,t);7.32(1H,t);7.62(1H,d);7.92(1H,d)

MS (ESP⁺) m/z 448 (M+H)⁺.

Anal.Calcd for C₂₂H₂₉N₃S₂O₃ 2.7HCl 0.3Et₂O C,49.0;H,6.15;N,7.39.

Found C,49.1;H,6.1;N,7.2.

Compound 31a

3-Nitro-1-napthoic acid 31a was synthesised from 3-nitro-1,8-napthalicanhydride according to the method of G. J. Leuck et al (Journal of theAmerican Chemical Society 51, 1831, 1929).

Compound 31b

3-Nitro-1-Napthoic acid 31a (5.0 g, 23.04 mmol) was coupled withL-Methionine methylester hydrochloride (analogously as for theequivalent step in Example 22) to give 31b as a white cyystalline solid.2.53 g (30.3%)

¹H NMR (CDCl₃,200 MHz) d2.0-2.5(5H,m);2.55-2.75(2H,m);3.85(3H,s);5.05(1H,m);6.9(1H,d,NH);7.6-7.85(2H,m);8.0-8.15(1H,m);8.3-8.5(2H,m);8.83(1H,m)

MS (ESP⁺) m/z 363 (M+H)⁺.

Compound 31c

31b (2.3 g, 6.35 mmol) was reduced (analogously as for the equivalentstep in Example 22) to give the corresponding aniline 31c as a yellowgum. 1.75 g (83%)

¹H NMR (CDCl₃,250 MHz) d2.05-2.2(4H,m);2.25-2.45(1H,m).2.63(2H,m);3.83(3H,s);5.03(1H,m);6.66(1H,d);7.05(1H,m);7.15(1H,m);7.28(1H,m);7.39(1H,m); 7.6(1H,m);8.15(1H,m)

MS (ESP+) m/z 333 (M+H)⁺,170.

Compound 31d

31c(1.7 g, 5.12 mmol) was coupled with the aldehyde 22b(1.76 g, 5.59mmol), (analogously as for the equivalent step in Example 30) to give31d as an off-white foam. 2.95 g(91.3%).

¹H NMR (CDCl₃+CD₃COOD,250 MHz) d1.5(9H,s),1.9(1H,m); 2.0-2.25(4H+CH₃COOH,m);2.25-2.44(1H,m);2.55-2.75(3Hm);3.25-3.53(2H,m);3.55-3.7(1H,m);3.7-3.95(4H,m);4.1-4.25(1H,m);4.25-4.4(1H,m);4.55-4.8(2H,m);5.03(1H,m);5.15-5.45(2H,m);5.96(1H,m);6.9-7.5(4H+CHCl₃,m);7.66(1H,m);8.1(1H,m)

MS (ESP+) m/z 632 (M+H)⁺.

Compound 31e

31d (2.0 g, 3.17 mmol) was deprotected (analogously as for theequivalent step in Example 15) to give the desired starting material 31eas a pale yellow foam. 1.62 g(93.4%).

¹H NMR (CDCl₃,300 MHz) d2.4-2.6(10H,m);1.85(4H,bs);2.0-2.2(4H,m);2.35(1H,m);2.5(1H,m);2.65(2H,t);2.9(1H,m);3.1(1H,m);3.3(1H,m);3.4(1H,m);3.55(1H,m);3.65(1H,m);3.8(3H,s);5.02(1H,m);6.65(1H,d);6.9(1H,m);7.1(1H,m); 7.2-7.3(1H+CHCl₃,m);7.4(1H,m);7.62(1H,m);8.1(1H,m)

MS (ESP+) m/z 548 (M+H)⁺, 448.

b) Compound 31f

31e(180 mg, 0.33 mmol) was hydrolysed (analogously as for Example 16)then purified by reverse phase HPLC (Dynamax® 60A,C₁₈, 8 m prep column),eluting with 50% MeOH/H₂O (0.1% TFA) to give product 31f as a whitefoam, 126 mg(65.9%).

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-1.8(1H,m);1.9-2.1(5H,m);2.4-2.7(3H+DMSO,m);3.0-3.1(1H,m);3.4-3.7(4H,m);3.75-3.9(1H,m);4.57(1H,m);6.9(1H,m);7.07(1H,m);7.17(1H,m);7.35(1H,m);7.63(1H,m);7.95(1H,m)

MS (ESP+) m/z 434 (M+H)⁺, 285.

Anal.Calcd for C₂₁H₂₇N₃S₂O₃.1.3TFA C,48.7;H,4.9;N,7.22.

Found C,48.6;H,4.9;N,7.1.

EXAMPLE 30

(See Scheme 37)

Preparation of a)(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester (compound 32) and; b)(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid (compound 32f) a) Preparation of Compound 32

Starting material compound 32e (55 mg, 0.096 mmol) was deprotected(analogously as for the equivalent step in Example 15) to give the titlecompound 32 as a white foam (56mg).

¹H NMR (CDCl₃,250 MHz) d1.6-1.85(1H,m);1.9-2.4(6H+CH₃C₅H₆);2.45-2.7(3H,m);3.1-3.25(1H,m);3.35-4.1(11H+H₂O,m);4.75-4.95(1H,m);6.8(1H,m);6.9-7.05(1H,m); 7.1-7.55(6H+CH₃C₆H₅+CHCl₃,m,)

MS (ESP+) m/z 474 (M+H)⁺.

Anal.Calcd for C₂₄H₃₁N₃O₃S₂. 2TFA.0.75toluene C,51.8;H,5.1;N,5.45.

Found C,51.6;H,5.2;N,5.1.

Starting material 32e was prepared as follows.

Compound 32a

Saturated NaHCO₃(aq) (90 mL) was added to a stirred solution ofmethyl-3-bromo-5-nitro-benzoate (4.0 g, 15.38 mmol) (Mindl and Vecera,Coll.Czech.Chem.Comm. 38.3496.1973.) and phenyl boronic acid (2.0 g,16.38 mmol) in dimethoxyethane (180 mL).Tetrakis(triphenylphosphine)palladium(0), (444 mg, 0.38 mmol) was addedand the mixture heated at reflux for 1 hr. The resulting black solutionwas allowed to cool to RT then quenched with saturated NaHCO₃(aq)(400mL). The aqueous was extracted with EtOAc(200 mL), then acidified to pH3with 2N HCl. The resulting suspension was filtered, washed with waterand azeotroped with toluene (3×25 mL) to give 32a as an off-white solidwhich was triturated with i-Hexane, filtered and dried, 2.6 g(69.5%).

¹H NMR (DMSO-D₆,300 MHz) d7.5(3H,m);7.8(2H,m);8.4-8.7(3H,m)

MS (ESP⁻) m/z 242 (M−H)⁻.

Anal. Calcd for C₁₃H₉NO₄: C,64.2;H,3.73;N,5.76.

Found C,64.0;H,3.7;N,5.6.

Compound 32b

32a (3.1 g, 12.76 mmol) was coupled with L-Methionine methylesterhydrochloride (analogously as for the equivalent step in Example 22) togive 32b, 4.9 g(99%).

¹H NMR (CDCl₃,200 MHz) d2.1-2.45(5H,m);2.65(2H,t);3.83(3H,s);4.99(1H,m);7.2-7.35(1H+CHCl₃,m,);7.4-7.6(3H,m);7.6-7.7(2H,m);8.38(1H,m) 8.58(2H,m)

MS (ESP+) m/z 389 (M+H)⁺.

Anal. Calcd for C₁₉H₂₀N₂O₅S C,58.8;H,5.19;N,7.21.

Found C,58.8;H,5.1;N,7.2.

Compound 32c

32b(3.0 g, 7.73 mmol) was reduced (analogously as for the equivalentstep in Example 30) to give the corresponding aniline 32c. 2.43g(87.8%).

¹H NMR (CDCl₃,250 MHz) d2.0-2.2(4H,m);2.2-2.4(1H,m);2.6(2H,m);3.8(3H,s); 3.9(2H,bs,NH₂);4.93(1H,m);6.93(1H,d,NHCO);7.03(1H,m);7.12(1H,m);7.3-7.5(4H,m);7.5-7.65(2H,m)

MS (ESP+) m/z 359 (M+H)⁺.

Compound 32d

32c (1.0 g, 2.8 mmol) was coupled with the aldehyde 22b (880 mg, 2.8mmol) (analogously as for the equivalent step in Example 30) to give32d. 1.51 g(82.3%)

¹H NMR (CDCl₃+CD₃COOD,250 MHz) d1.5(9H,s);1.8-2.0(1H,m); 2.0-2.4(5H+CH₃COOH,m);2.5-2.75(3H,m);3.2-3.45(2H,m);3.5-3.7(1H,m);3.7-3.9(4H,m);4.0-4.4(2H,m);4.5-4.75(2H,m);4.9-5.05(1H,m);5.1-5.45(2H,m)5.8-6.1(1H,m);7.03(1H,m);7.1-7.5(5H+CHCl₃,m);7.55-7.7(2H,m)

MS (ESP+) m/z 658 (M+H)⁺.

Anal.Calcd for C₃₃H₄₃N₃O₇S₂0.1H₂O C,59.9;H,6.61;N,6.35.

Found C,59.7;H,6.8;N,6.2.

Compound 32e

32d(1.1 g, 1.67 mmol) was deprotected (analogously as for the equivalentstep in Example 15) to give the desired starting material 32e, 800mg(83.4%).

¹H NMR (CDCl₃,250 MHz) d1.25(1.5H,t,CH ₃CH₂COCH₃);1.4-1.6(10H,m);1.9(2H,bs,NH+H₂O);2.0-2.22(4H+CH₃CH₂CO₂CH ₃);2.23-2.55(2H,m);2.51-2.65(2H,m);2.9(1H,m);3.12(1H,m);3.2-3.75(4H,m);3.8(3H,m);4.13(1.3H,q,CH₃CH ₂CO₂CH₃);4.45(1H,bs,NH);4.95(1H,m);6.85-7.0(2H,m,ArH+NHCO);7.07(1H,m);7.2-7.5(4H+CHCl3,m);7.5-7.65(2H,m)

MS (ESP+) m/z 574 (M+H)⁺,474.

Anal.Calcd for C₂₉H₃₉N₃O₅S₂ 0.5EtOAc C,60.3;H,7.02;N,6.8.

Found C,59.9;H,7.1;N,6.6.

b) Preparation of Compound 32f

Starting material 32e (140 mg, 0.244 mmol) was hydrolysed (analogouslyas for the equivalent step in Example 31) to give the desired product32f as a white foam, 96.3 mg (64.9%).

¹H NMR (DMSO-D₆+CD₃COOD,250 MHz) d 1.5-1.8(1H,m);1.9-2.2(5H,m);3.05(1H,q);3.15-3.6(7H,m);3.65-3.9(1H,m);4.45-4.65(1H,m);6.95-7.05(1H,m);7.05-7.2(1H,m);7.25-7.5(4H,m);7.55-7.7(2H,m).

MS (ESP+) m/z 460 (M+H)⁺,279.

Anal.Calcd for C₂₃H₂₉N₃S₂O₃ 1.3TFA C,50.6;H,5.02;N,6.91.

Found C,50.6;H,5.1;N,7.2.

The starting material was prepared as described in a) immediately above.

EXAMPLE 31

(See Scheme 38)

Preparation of a)(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester (compound 33) and; b)(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid (compound 33f) a) Preparation of Compound 33

Starting material 33e (53.4 mg, 0.093 mmol) was deprotected (analogouslyas for the equivalent step in Example 31) to give the title compound 33as a white solid, 43.2 mg(87%).

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-1.9(3H+CH ₃COOH,m);1.95(3H,s);2.0-2.3(2H,m);2.4-2.65(1H+DMSO,m);3.0-3.15(1H,m);3.3-3.9(8H,m);4.25-4.4(1H,m);6.7(1H,m);6.78(1H,m); 7.1-7.4(6H,m).

MS (CI⁺) m/z 474 (M+H)⁺.

Anal.Calcd for C₂₄H₃₁N₃S₂O₃ 1.75TFA C,53.6;H,6.14;N,7.82.

Found C,53.6;H,6.3;N,7.7.

The starting material was prepared as follows.

Compound 33a

2-Bromo-5-nitrobenzoic acid (12.28 g, 0.05 mmol) was coupled withbenzene boronic acid (6.7 g, 0.055 mmol), (analogously as for theequivalent step in Example 32) to give 33a as a white solid, 10.95g(90.3%).

¹H NMR (DMSO-D₆,300 MHz)d7.3-7.5(5H,m);7.65(1H,m);8.35(1H,m);8.45(1H,m).

MS (ESP−) m/z 242 (M−H)⁻,198.

Compound 33b

33a (3.58 g, 14.7 mmol) was coupled with L-Methionine methyl esterhydrochloride (3.25, 16.2 mmol), (analogously as for the equivalent stepin Example 32) to give 33b as a pale yellow solid, 3.02 g(52.6%)

¹H NMR (CDCl₃,300 MHz) d1.7-2.2(7H,m);3.7(3H,s);4.7(1H,m);6.05(1H,m,NH);7.35-7.6(6H,m)8.33(1H,m);8.55(1H,m)

MS (ESP+) m/z 389 (M+H)⁺.

Compound 33c

33b(1.0 g, 2.6 mmol) was reduced (analogously as for the equivalent stepin Example 30) to give the corresponding aniline 33c, 725 mg(78.6%).

¹H NMR (CDCl₃,300 MHz) d1.6-1.8(1H,m);1.8-2.15(6H,m);3.6(3H,s);3.7-3.9(2H,bs,NH ₂);4.6-4.7(1H,m);5.85(1H,d,NHCO);6.79(1H,m);7.0(1H,m);7.15(1H,d);7.2-7.45(5H+CHCl₃,m).

MS (ESP+) m/z 359,(M+H)⁺,196.

Compound 33d

33c (710 mg, 1.98 mmol) was coupled with the aldehyde 22b (625 mg, 1.98mmol) (analogously as for the equivalent step in Example 30) to give33d, 1.1 g(84.4%).

¹H NMR (CDCl₃+CD₃COOD,250 MHz) d1.5(9H,s);1.6-2.2(8H+CH ₃COOH,m);2.5-2.75(1H,m),3.2-3.4(2H,m);3.45-3.9(5H,m);4.05-4.35(2H,m);4.5-4.8(3H,m);5.15-5.45(2H,m);5.8-6.1(1H,m);6.75-6.9(1H,m);6.9-7.05(1H,m);7.1-7.23(1H,m);7.25-7.45(5H+CHCl₃,m).

MS (ESP+) m/z 658 (M+H)⁺.

Anal.Calcd for C₃₃H₄₃N₃S₂O₇ C,60.3;H,6.59;N,6.39.

Found C,60.0;H,6.9;N,6.2.

Compound 33e

33d (1.0 g, 1.52 mmol) was deprotected (analogously as for theequivalent step in example 15) to give the desired starting material33e, 658 mg(75.4%).

¹H NMR (CDCl₃+CD₃COOD,250 MHz) d1.5(9H,s);1.6-2.2(8H+CH ₃COOH,m);2.55-2.75(1H,m);3.25-3.4(1H,m);3.5-3.75(5H,m);3.75-4.2(3H,m);4.55-4.75(1H,m);6.7-6.85(1H,m);6.85-6.97(1H,m);7.1-7.25(1H,m);7.25-7.48(5H+CHCl₃,m).

MS (ESP+) m/z 574 (M+H)⁺,474.

Anal.Calcd for C₂₉H₃₉N₃O₅S₂ C,60.7;H,6.85;N,77.32.

Found C,60.7;H,7.20;N,7.30.

b) Preparation of Compound 33f

Starting material 33e (100 mg, 0.174 mmol) was hydrolysed (analogouslyas for the equivalent step in Example 31) to give 33f as a white foam,64.6 mg(59.8%).

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-2.0(6H+CH ₃COOH,m);2.0-2.3(2H,m);2.3-2.7(1H+DMSO);3.0-3.1(1H,m);3.2-3.9(5H,m);4.2-4.35(1H,m);6.6-6.9(2H,m);7.1-7.4(6H,m).

MS (ESP+) m/z 460 (M+H)⁺,311.

Anal Calcd for C₂₃H₂₉N₃O₃S₂ 1.4TFA C,50.0;H,4.95;N,6.79.

Found C,49.9;H,5.1;N,6.7.

Starting material 33e was prepared as described in a) immediately above.

EXAMPLE 32

(See Scheme 39)

Preparation of a)(2S)-2-{2-Benzyl-5-[(4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid methyl ester (compound 34) and; b)(2S)-2-{2-Benzyl-5-[(4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid (compound 34h) a) Preparation of Compound 34

Starting material 34g (500 mg, 0.85 mmol) was deprotected (analogouslyas for the equivalent step in Example 31) to give the title compound 34as a white solid, 454 mg (89.3%).

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-1.7(1H,m);1.85-2.1(5H,m);2.35-2.6(3H+DMSO,m);2.9-3.1(1H,m);3.1-3.8(8H,m);3.9(2H,q);4.4-4.6(1H,m);6.5-6.7(>1H,m);6.9-7.0(1H,m);7.0-7.3(6H,m).

MS (ESP+) m/z 488 (M+H)⁺,325.

Anal.Calcd for C₂₅H₃₃N₃S₂O₃.3HCl C,50.3;H,6.08;N,7.04.

Found C,50.4;H,6.3;N,7.3.

Starting material 34g was prepared as follows.

Compound 34a

A solution of 2-bromo-5-nitrobenzoic acid (9.0 g, 36.6 mmol) in MeOH(200 mL) was treated with SO₂Cl₂(2.0 mL) and the resulting solutionheated at reflux for 18 hrs. The reaction mixture was then evaporated,pre-absorbed on SiO₂ (Merck, 9385) and chromatographed, eluting with 10%EtOAc/i-Hexane. Appropriate fractions were combined and evaporated togive 34a as a crystalline white solid, 8.38 g(88.1%)

¹H NMR (CDCl₃,300 MHz)d4.0(3H,s,CO₂CH₃);7.85(1H,m);8.18(1H,m);8.63(1H,m).

Compound 34b

A solution of benzyl bromide (2.0 mL, 17.3 mmol) in THF(10 mL) was addeddropwise at 0° C. to a stirred suspension of zinc dust(1.7 g, 26 mmol)in THF(10 mL) which had been activated according to the method describedby Knochel (J.O.C. 53,2392,1988). The mixture was left to warm to RT andstir for 3 hrs. An aliquot (6.5 mmol) of the supernatent containing thebenzyl zinc reagent was then added to a stirred solution of 34a (1.0 g,3.85mmol) and Pd(PPh₃)₂Cl₂ (27 mg, 0.0385 mmol) in THF(10 mL) at RTunder argon. After 1 hr a second aliquot (6.5 mmol) of the benzyl zincreagent was added. The resulting black reaction mixture was quenchedwith 2N HCl (250 mL) and extracted with EtOAc (2×100 mL). The combinedorganics were washed with water (50 mL) and brine (50 mL), filteredthrough phase separating paper and evaporated to an orange gum. This waschromatographed on SiO₂ (Merck, 9385) eluting with 10% EtOAc/i-Hexane togive 34b as a yellow oil, 590 mg(56.6%).

¹H NMR (CDCl₃,300 MHz) d3.9(3H,s,CO2CH ₃);4.48(2H,s,CH₂Ph);7.0-7.5(6H,m); 8.23(1H,m);8.75(1H,m).

MS (ESP⁻) m/z 270 (M−H)⁻,210.

Compound 34c

2N NaOH (2.0 mL, 4 mmol) was added to a solution of 34b (560 mg, 2.06mmol) in MeOH (10 mL) at RT. After 2 hrs the RM was evaporated to removethe MeOH and then partitioned between Et₂O (20 mL) and 2N NaOH (20 mL).The aqueous was acidified to pH2/3 with 2N HCl and extracted withEtOAc(3×20 mL). The combined organics were washed with water (20 mL) andbrine (20 mL), filtered through phase separating paper and evaporated toyield 34c as a white solid, 453 mg(85.3%).

¹H NMR (DMSO-D₆,300 MHz) d4.45(2H,s,CH ₂Ph);7.0-7.4(5H,m);7.55(1H,m);8.3(1H,m);8.53(1H,m).

MS (ESP⁻) m/z 256 (M−H)⁻,212.

Compound 34d

34c (630 mg, 2.45 mmol) was coupled with L-Methionine methyl esterhydrochloride (540 mg, 2.7 mmol), (analogously as for the equivalentstep in Example 32) to give 34d as a pale yellow solid, 900 mg (91.3%).

¹H NMR (DMSO-D₆,250 MHz) d1.9-2.25(5H,m);2.5-2.75(2H+DMSO,m);3.74(3H,s,CO2CH3);4.28(2H,q,CH ₂Ph);4.55-4.75(1H,m);7.15-7.5(5H,m);7.6(1H,m);8.2-8.35(2H,m);9.13(1H,d,NHCO).

MS (ESP+) m/z 403 (M+H)⁺.

Compound 34e

SnCl₂2H₂O (2.5 g, 11.08 mmol) was added to a stirred solution of 34d(900 mg, 2.24 mmol) in EtOAc(50 mL) and the resulting mixture heated atreflux for 18 hrs. The RM was cooled to RT and treated with 0.88S0 SGNH₃(aq) dropwise to pH8. The resulting heavy white precipitate wasremoved by filtration through celite(545). The filtrates were thenevaporated and purified by chromatography (Mega Bond Elut, SiO₂),eluting with CH₂Cl₂ and then 50% EtOAc/i-Hexane to give thecorresponding aniline 34e, 595 mg(71.4%).

¹H NMR (CDCl₃,300 MHz) d1.75-2.2(5H,m);2.25-2.45(2H,m);3.6-3.8(5H,m,CO2CH ₃+NH ₂);4.08(2H,q,CH₂Ph);4.65-4.85(1H,m);6.24(1H,d,NHCO);6.7(1H,m);6.78(1H,m);7.0(1H,m);7.05-7.3(5H+CHCl₃,m).

MS (ESP+) m/z 373 (M+H)⁺,210.

Compound 34f

34e (570 mg, 1.53 mmol) was coupled with the aldehyde 22b (580 mg, 1.84mmol) (analogously as for the equivalent step in Example 30) to give 34fas a crude pale green foam(1.54 g).

MS (ESP+) m/z 672 (M+H)⁺.

Compound 34g

34f (1.5 g, 2.24 mmol) was deprotected (analogously as for theequivalent step in Example 15) to give the desired starting material 34gas a pale brown glass, 550 mg (41.9%).

¹H NMR (CDCl₃,300 MHz) d1.3-1.65(10H,m);1.7-2.2(5H+H₂O,m);2.25-2.6(3H,m);2.8-3.9(9H,m);3.9-4.25(2H,m);4.6-4.9(1H,m);6.3(1H,d,NHCO);6.55-6.8(2H,m);6.9-7.4(5H+CHCl₃,m).

MS (ESP+) m/z 588 (M+H)⁺,488.

b) Preparation of Compound 34h

Starting material 34g (52 mg, 0.087mmol) was hydrolysed (analogously asfor the equivalent step in Example 16), then purified by reverse phaseHPLC (Dynamax® 60A, C₁₈, 8 m prep column), eluting with 50% MeOH/H₂O(0.% TFA) to give 34h as a colourless glass, 38.2 mg(56.6%).

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-1.7(1H,m);1.8-2.1(5H+CH₃COOH,m);2.3-2.6(3H+DMSO,m);2.9-3.1(1H,m);3.2-4.1(7H,m);4.3-4.5(1H,m);6.5-6.7(2H,m);6.9-7.0(1H,m);7.05-7.25(5H,m).

MS (ESP+) 474 (M+H)⁺.

Anal.Calcd for C₂₄H₃₁N₃S₂O₃ 1.4TFA C,50.8;H,5.16;N,6.14.

Found C,51.0;H,5.3;H,6.7.

The starting material was prepared as described in a) immediately above

EXAMPLE 33

(See Scheme 40)

Preparation of a)(2S)2-{2-Benzyl-4-[([2S,4S]4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid methyl ester (compound 35) and; b)(2S)2-{2-Benzyl-4-[([2S,4S]4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid (compound 35 g) a) Preparation of Compound 35

The title compound 35 was synthesised frommethyl-2-bromo-4-nitro-benzoate using the same methodology as describedin Example 32 but using Pd₂(dba)₃ as a source of catalytic palladium inthe benzylation reaction.

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-1.7(1H,m);1.8-2.1(5H,m);2.3-2.6(3H+DMSO,m);2.9-3.1(1H,m);3.2-3.8(8H,m);4.05(2H,m);4.44-4.6(1H,m);6.4-6.6(2H,m);7.0-7.35(6H,m)

MS (ESP+) m/z 488(M+H)⁺,325.

Anal Calcd for C₂₅H₃₃N₃S₂O₃.2HCl C,53.6;H,6.29;N,7.5.

Found C,53.5;H,6.5;N,7.3.

b) Preparation of Compound 35 g

Compound 35 (100 mg,0.18 mmol; see a) above) was hydrolysed (analogouslyas for the equivalent step in Example 32) to give 35 g as a white solid,85.8 mg(67.3%).

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.5-1.7(1H,m);1.8-2.1(5H,m);2.3-2.6(3H+DMSO,m);2.9-3.9(6H,m);3.95-4.2(2H,m);4.3-4.6(1H,m);6.4-6.5(2H,m);7.0-7.3(6H,m)

MS (ESP+) m/z 474(M+H)⁺,325.

Anal Calcd for C₂₄H₃₁N₃S₂O₃.1.3TFA C,51.4;H,5.24;N,6.76.

Found C,51.2;H,5.4;N,6.7.

EXAMPLE 34

(See Scheme 41)

(2S)2-{2Benzyl-5-[([2S,4S]4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid isopropyl ester (compound 36)

The nitro compound 36b was reduced to the corresponding aniline, coupledwith the thioproline aldehyde 22b using IPA as solvent and deprotectedexactly analogously as for Example 32 to give the title compound 36.

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.0-1.3(6H,m);1.5-1.7(1H,m);1.8-2.1(5H,m);2.3-2.6(3H+DMSO,m);2.9-4.1(8H,m);4.3-4.6(1H,m);4.8-5.0(1H,m);6.5-6.7(2H,m);6.8-7.3(6H,m)

MS (ESP+) m/z 516(M+H)³⁰,325.

Anal Calcd for C₂₇H₃₇N₃S₂O₃.2HCl C,55.1;H,6.68;N,7.14.

Found C,54.9;H,7.0;N,7.1.

Compound 36a

A solution of 34d (25.24 g, 62.78 mmol) in MeOH (500 mL) was treatedwith 2N NaOH (35 mL, 70 mmol). The resulting solution was thenevaporated to dryness and the solids partitioned between Et₂O (200 mL)and water (500 mL). The aqueous was then acidified to pH2 with 2N HCland extracted with EtOAc(2×250 mL). The combined organics were washedwith water(2×100 mL), brine(100 mL), filtered through phase separatingpaper and evaporated to give 36a as a white solid, 23.57 g(96.8%).

¹H NMR (DMSO-D₆,300 MHz) d1.8-2.2(5H,m);2.3-2.6(2H+DMSO,m);4.1-4.3(2H,m);4.4-4.6(1H,m);7.1-7.3(5H,m);7.4-7.6(1H,m);8.1-8.3(2H,m);8.9-9.0(1H,m,NHCO)

MS (ESP−) m/z 387(M-H)⁻.

Compound 36b

Sulphuryl chloride (5.0 mL, 62 mmol) was added to a stirred suspensionof 36a (19.2 g, 50 mmol) in IPA (500 mL). The resulting mixture was thenheated at reflux for 18 hrs. The reaction mixture was then evaporated to⅕ volume and partitioned between EtOAc (1 L) and saturated NaHCO₃ (aq)(500 mL). The organics were then washed with water (200 mL), brine (200mL), filtered through phase separating paper and evaporated to give 36bas a white solid, 21.25 g(100%)

¹H NMR (DMSO-D₆,.300 MHz) d1.0-1.3(6H,m);1.8-2.2(5H,m);2.3-2.6(2H+DMSO,m);4.1-4.3(2H,m);4.4-4.6(1H,m);4.8-5.0(1H,m);7.1-7.3(5H,m);7.4-7.6(1H,m);8.1-8.3(2H,m);9.0(1H,m,NHCO)

MS (ESP+) m/z 431(M+H)⁺.

EXAMPLE 35

(See Scheme 42)

(2S)2-{2-Benzyl-5-[N-([2S,4S]4-sulfanylpyrrolidin-2-ylmethyl)-N-(3-methoxypropionyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid isopropyl ester (compound 37)

Starting material 37b was deprotected using the same methodology for theequivalent step described in Example 32 to give the title compound 37.

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) d1.0-1.3(6H,m);1.5-1.7(1H,m);1.8-2.1(5H,m);2.2-2.6(5H+DMSO,m);2.9-3.95(10H,m);4.0-4.2(3H,m),4.4-4.6(1H,m);4.8-5.0(1H,m);7.0-7.5(8H,m)

MS (ESP+) m/z 602 (M+H)⁺.

Anal Calcd for C₃₁H₄₃N₃S₂O₅.1.5HCl C,56.7;H,6.83;N,6.4.

Found C,56.7;H,7.0;N,6.0.

The starting material was prepared as follows.

EEDQ (530 mg,2.15 mmol) was added to a stirred solution of 36d (1.5 g,2.15 mmol; see Example 34) and 3-methoxy propionic acid (220 mL, 2.36mmol) in CH₂Cl₂ (15 mL). The mixture was left to stir 18 hrs at RT thenevaporated. The residues were then partitioned between 1N citricacid(aq) (200 mL) and EtOAc (100 mL). The organics were washed withsaturated NaHCO₃ (aq) (50 mL), water(50 mL) and brine(50mL), filteredthrough phase separating paper and evaporated to give a pale yellow gum.This was then purified by flash chromatography on SiO₂ (Merck 9385)eluting a gradient of 0-50% EtOAc/i-Hexane. Appropriate fractions werefiltered and evaporated to give starting material 37b as a colourlessgum,1.14 g(67.7%).

MS (ESP+) m/z 786 (M+H)⁺.

EXAMPLE 36

(See Scheme 43)

Preparation of a)N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-1-yl-ethyl)-butyramide(compound 56) and; b)N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-N-(2-naphthalen-1-yl-ethyl)-2-pyridin-3-yl-acetamide(compound 57) a) Preparation of compound 56

The method described in Example 23 for the synthesis of compound (6) wasused to prepare compound (56) as set out in Scheme 43.

NMR data in CDCl₃ δ0.91(s, 9H), 1.5(m, 1H), 1.75(m, 1H), 1.82(d,1H),1.91(d, 1H), 2.52(m, 1H), 2.92(m, 1H), 3.33(m, 3H), 3.72(m, 4H), 4.15(m,1H), 7.26(d,1H), 7.41(t, 1H), 7.56(m, 2H), 7.8(d, 1H), 7.9(2d, 2H),9.08(br,s, 1H).

Micro Analysis: %Theory C,64.2; H,7.97; N,6.5. (1.00 HCl, 0.5H₂O %FoundC,64.4; H,7.90; N,6.3.

Starting material compound (54) was synthesised analogously with Example23 using the appropriate intermediates:

Compound (52).

NMR data in CDCl₃ δ1.00(2s, 9H), 1.46(d, 9H), 1.95(m, 2H), 2.4(m, 2H),3.3(m, 4H), 3.7(m, 3H), 4.00(m, 3H), 4.57(d, 2H), 5.22(2d, 2H), 5.90(m,1H), 7.24-8.4(m, 7H).

Compound (54).

NMR data in CDCl₃ δ1.00(2s, 9H), 1.35(m, 1H), 1.49(s, 9H), 1.89(br,s,1H), 1.95(d, 1H), 2.3(m, 1H), 2.32(d, 1H), 2.88(2q, 1H), 3.1-3.9(m, 9H),7.25-8.31(m, 7H).

b) Preparation of Compound 57

The method described in Example 24 for the synthesis of compound (27)was used in a similar manner to prepare compound (57).

NMR data in CDCl₃ δ1.2(m, 1H), 2.00(m, 1H), 2.6(m, 2H), 3.15-4.40(m,10H), 7.28-8.70(m, 11H), 9.4(br,s, 1H).

Micro Analysis: %Theory C,56.0; H,6.20; N,8.17. (2HCl, 2H₂O) %FoundC,56.4; H,6.46; N,7.70.

Starting material compound (55) was synthesised analogously with Example24 using appropriate intermediates:

Compound (53).

NMR data in CDCl₃ δ1.48(s, 9H), 1.84(m, 1H), 2.42(m, 1H), 2.87-3.45(m,5H), 3.63-4.26(m, 7H), 4.55(d, 2H), 5.22(2d, 2H), 5.9(m, 1H), 7.1-8.7(m,11H).

Compound (55),

NMR data in CDCl₃ δ1.34(m, 1H), 1.5(s, 9H), 1.95(m, 1H), 2.32(m, 2H),2.72-4.00(m, 10H), 7.1-8.6(m, 11H).

Example 37

(See Scheme 44)

Preparation of a)N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-butyramide(compound 67); b)N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-butyramide(compound 68); c)N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-2-pyridin-3-yl-acetamide(compound 69) and; d)N-(2,2-Diphenyl-ethyl)-1-oxy-N-([2S,4S]-sulfanyl-pyrrolidin-2-ylmethyl)-6-methoxy-nicotinamide(compound 70). a) Preparation of Compound 67

The method described in Example 23 for the synthesis of compound (6) wasused in an analogous manner to prepare compound (67) using appropriateintermediates—see Scheme 44.

NMR data in DMSO-d6 δ0.75(m, 6H), 1.55(m,1H), 1.87(m, 2H), 2.05-2.45(m,1H), 3.05(m,1H), 3.25-3.70(m, 6H), 4.05(m,2H), 4.20-4.55(m,1H),7.30(m,10H), 8.80-9.80(2br,s, 2H)

Micro Analysis: %Theory C,63.9; H,7.82; N,6.21. (1.00HCl,1.00H₂O) %FoundC,64.1; H,7.70; N,6.00.

Compound (58).

NMR data in CDCl₃ δ1.50,(s, 9H), 1.77(m,1H), 2.40(m,1H), 2.75(m,1H),3.00(m,1H), 3.14(q,1H), 3.24(d, 2H), 3.67(m,1H), 3.93(m,1H), 4.10(m,2H), 4.54(d, 2H), 5.25(m, 2H), 5.90(m,1H), 7.25(m,10H)

Compound (59).

NMR data in CDCl₃ δ0.85(m, 6H), 1.48(m, 9H), 1.80(m, 2H), 2.10(m, 2H),2.40(m,1H), 2.80-4.35(m, 9H), 4.55(m, 2H), 5.25(m, 2H), 5.90(m,1H),7.25(m,10H)

Compound (63).

NMR data in CDCl₃ δ0.85(2d, 6H), 1.24(m,1H), 1.48(s, 9H), 1.68(m,1H),1.81(d,1H), 1.95-2.35(m, 3H), 2.75-3.65(m, 6H), 3.90-4.55(m, 3H),7.25(m,10H)

b) Preparation of Compound 68

Similarly compound (68) was synthesised from compound (60) as set out inScheme 44.

Compound (68)

NMR data in DMSO-d6 δ0.85(m, 9H), 1.55(m,1H), 1.74-2.27(m,2H),2.37(m,1H), 3.05(m,1H), 3.45(m, 6H), 4.05(m, 2H), 4.18-4.55(m,1H),7.28(m,10H), 8.90-9.90(m, 2H)

Micro Analysis: %Theory C,64.6; H,8.02; N,6.02. (1.0HCl,1.0H₂O) %FoundC,64.8; H,8.30; N,5.70.

Compound (60).

NMR data in CDCl₃ δ0.93(m, 9H), 1.50(s, 9H), 1.82(m, 2H), 2.35(m, 3H),2.90-4.35(m, 8H), 4.55(m, 2H), 5.25(m,2H), 5.90(m,1H), 7.25(m,10H).

Compound (64).

NMR data in CDCl₃) δ0.93(s, 9H), 1.24(m,1H), 1.48(s, 9H), 1.80(q,1H),2.23(d,1H), 2.30(m,1H), 2.75-3.70(m, 6H), 3.90-4.60(m, 3H), 7.25(m,10H).

c) Preparation of Compound 69

Compound (69) was synthesised from compound (61) (see Scheme 44)analogously with the procedure described in Example 24 for thepreparation of compound (27).

Compound (69).

NMR data in CDCl₃ δ1.95(m,1H), 2.40(m,1H), 2.60(m,1H), 3.15-4.50(m,11H),7.28(m,10H), 7.67(m,1H), 8.05(m,1H), 8.50(m,1H), 8.71(m,1H),9.10-10.20(br,d, 2H).

Micro Analysis: %Theory C,55.1; H,5.51; N,7.01. (2.0 HCl,0.75TFA,0.5H₂O)%Found C,55.0; H,5.60; N,6.90.

Compound (61).

NMR data in CDCl₃ δ1.47(s, 9H), 1.80(m,1H), 2.30-4.65(m,14H), 5.23(m,2H), 5.90(m,1H), 7.25(m,12H), 8.10-8.55(m, 2H).

Compound (65).

NMR data in CDCl₃ δ1.25(m,1H), 1.48(s, 9H), 2.30(m,1H),2.70-4.55(m,12H), 7.30(m,12H), 8.28(2d,1H), 8.45(m,1H).

d) Preparation of Compound 70

Similarly compound (70) was synthesised from compound (62) usingappropriate intermediates.

NMR data in CDCl₃ δ1.67(m,1H), 2.15(d,1H), 2.47(m,1H), 3.16(br,s, 1H),3.50(m, 2H), 3.85-4.40(m, 8H), 5.22(br,s,1H), 6.56(d,1H),7.00-7.35(m,11H), 7.90(s,1H), 8.85-10.75(2br,s, 2H)

Micro Analysis %Theory C,57.2; H,5.91; N,7.70. (2.0HCl,0.5H₂O) %FoundC,57.5; H,5.60; N,7.30.

Compound (62).

NMR data in CDCl₃ δ1.50(s, 9H), 1.60(m,1H), 2.47(m,1H),3.00-4.50(m,12H), 4.58(d, 2H), 5.25(m, 2H), 5.90(m,1H), 6.53(d,1H),6.95(m,1H), 7.25(m,11H).

Compound (66).

NMR data in CDCl₃ δ1.20(m,1H), 1.45(s, 9H), 2.30(m,1H), 2.66(m,1H),3.00-3.45(m, 4H), 3.55(m,1H), 3.95-4.25(m, 5H), 4.47(m,1H), 6.55(d,1H),7.25(m,11H), 7.65(m,1H).

EXAMPLE 38

(See Scheme 45)

Preparation of a)N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-N-(2-naphthalen-2-yl-ethyl)-butyramide(compound 80); b)N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-2-yl-ethyl)-butyramide(compound 81); c)N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-N-(2-naphthalen-2-yl-ethyl)-2-pyridin-3-yl-acetamide(compound 82) and; d)N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-2-(4-methoxy-phenyl)-N-(2naphthalen-2-yl-ethyl)-acetamide(compound 83). a) Preparation of Compound 80

The method described in Example 23 for the synthesis of compound (6) wasused to prepare compound (80).

NMR data in DMSO-d6 δ0.75(m, 6H), 0.87(d,1H), 1.65(m,1H), 1.92(m,1H),2.02(d,1H), 3.03(m, 3H), 3.20-3.80(m, 9H), 7.48(m, 3H), 7.75(d,1H),7.85(m, 3H), 8.90-9.90 (br,d, 2H)

Micro Analysis: %Theory C,64.9; H,7.68; N,6.88. (1.00 HCl) %FoundC,64.9; H,7.50; N,6.80.

Starting material compound (76) was synthesised analogously with Example23 using appropriate intermediates—see Scheme 45.

Compound (71).

NMR data in CDCl₃ δ1.50(s, 9H), 1.85(m,1H), 2.50(m,1H), 2.80(m,1H),3.00(m, 5H), 3.20(m,1H), 3.65(m,1H), 4.00(m,1H), 4.10(m,1H), 4.53(d,2H), 5.20(m, 2H)5.90(m,1H), 7.32(m,1H), 7.42(m, 2H), 7.63(s,1H), 7.80(m,3H).

Compound (72).

NMR data in CDCl₃ δ0.90(m, 7H), 1.00-2.60(m, 14H), 3.00(m, 2H),3.10-4.20(m, 7H), 4.60(m, 2H), 5.25(m, 2H), 5.90(m,1H), 7.30-7.50(m,3H), 7.60(m,1H), 7.80(m, 3H).

Compound (76).

NMR data in CDCl₃ δ0.90(m, 6H), 1.10-2.50(m,15H), 2.80-3.80(m, 9H),7.26-7.50(m, 3H), 7.60(m,1H), 7.80(m, 3H)

b) Preparation of Compound 81

Compound (81) was synthesised from compound (73) as set out in Scheme 45in a similar manner to preparation of compound 80 (see above).

NMR data in DMSO-d6 δ1.08(d, 9H), 1.80(m,1H), 2.15(m, 2H), 2.65(m,1H),3.00-4.00(m,10H), 7.63(m, 3H), 7.90(s,1H), 8.03(m, 3H), 9.50(br,d, 2H).

Micro Analysis: %Theory C,64.9; H,7.93; N,6.58. (1.0HCl,0.25H₂O) %FoundC,64.8; H,8.10; N,6.50.

Compound (73).

NMR data in CDCl₃ δ1.00(m, 9H), 1.47(s, 9H), 1.80-2.55(m, 4H), 3.00(m,2H), 3.10-4.20(m, 8H), 4.60(d, 2H), 5.25(m, 2H), 5.90(m,1H),7.30-7.85(m, 7H)

Compound (77).

NMR data in DMSO-d6(100° C.) δ0.95(m, 9H), 1.35-1.75(m, 9H), 2.15(s,2H), 2.40(m,1H), 2.60-3.90(m,12H), 7.40(m, 3H), 7.70(m,1H), 7.80(m, 3H).

c) Preparation of Compound 82

Compound (82) was synthesised from compound (74) as set out in Scheme 45by a similar procedure to that described in Example 24 for thepreparation of compound (27).

Compound (82).

NMR data in DMSO-d6 δ1.65(m,1H), 2.90-4.15(m,14H), 7.35-8.90(m,11H),9.50(br,d, 2H).

Micro Analysis: %Theory C,51.9; H,5.19; N,6.99. (2.0HCl,1.0TFA,0.5H₂O)%Found C,52.2; H,5.40; N,7.00.

Compound (74).

NMR data in DMSO-d6 (100° C.) δ1.45-1.75(m,10H), 2.85-3.85(m,11H),4.03(m,1H), 4.20(m,1H), 4.45-4.65(m, 2H), 5.20(m, 2H), 5.90(m,1H),7.23(m,1H), 7.45(m, 4H), 7.67(s,1H), 7.80(m,3H), 8.35(m, 2H).

Compound (78)

NMR data in DMSO-d6 (100° C.) δ1.30-1.75(m, 9H), 2.40(m,1H),2.55-3.90(m,14H), 7.10-8.45(m, 11H).

d) Preparation of Compound 83

Similarly compound (83) was synthesised from compound (75) usingappropriate intermediates as set out in Scheme 45.

Compound (85).

NMR data in DMSO-d6 δ1.65(m,1H), 2.95(m, 2H), 3.08(m,1H),3.25-4.00(m,13H), 6.80(m, 2H), 7.06(2d, 2H), 7.47(m, 3H), 7.68(d,1H),7.85(m, 3H), 9.35(br,d, 2H).

Micro Analysis: %Theory C,62.7; H,6.57; N,5.62. (1.5 HCl,0.5H₂O) %FoundC,62.4; H,6.50; N,5.40.

Compound (75).

NMR data in DMSO-d6 (100° C.) δ1.45(s, 9H), 1.75(m,1H),2.75-3.85(m,14H), 4.00(m,1H), 4.14(m,1H), 4.45-4.65(m, 2H), 5.20(m, 2H),5.90(m,1H), 6.80(m, 2H), 7.05(m, 2H), 7.33(m,1H), 7.45(m, 2H),7.63(s,1H), 7.80(m, 3H).

Compound (79).

NMR data in DMSO-d6 (100° C.) δ1.30-1.75(m, 9H), 2.35(m,1H),2.60-3.90(m,17H), 6.78(m, 2H), 7.05(m, 2H), 7.40(m, 3H), 7.65(m,1H),7.80(m, 3H).

EXAMPLE 39

(See Scheme 46)

Preparation of a)(2S)-2-({2-phenyl-4-[((2S,4S)-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanyl-butyricacid methyl ester (compound 38) and; b)(2S)-2-({2-phenyl-4-[((2S,4S)-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanyl-butyricacid (compound 38f). a) Preparation of Compound 38

Methyl-2-bromo-4-nitro-benzoate was coupled with phenyl boronic acid(analogously as for the equivalent step in Example 30) then coupled anddeprotected using the same methodology as previously described forExample 32 to give the title compound 38.

¹H NMR (DMSO-D₆,250 MHz) δ1.35-1.75(3H,m);1.8(3H,s);1.9-2.2(2H,);2.25-2.5(2H+DMSO,m);2.75-3.9(10H,m);4.0-4.25(1H,m);5.0-5.9(5H,bs,H₂O);6.3-6.6(2H,m);7.0-7.3(7H,m);7.95(1H,m);9.2-9.8(2H,bd).

MS (ESP+) m/z 474 (M+H)⁺,311,196.

Anal.Calcd for C₂₄H₃₁N₃O₃S₂.2HCl.1.5H₂O C,50.3;H,6.3;N,7.3.

Found C,50.4;H,6.1;N,7.3.

b) Preparation of Compound 38f

Compound 38 was hydrolysed to the corresponding acid (analogously as forthe equivalent step in Example 33)o give 38f.

¹H NMR (DMSO-D₆+CD₃COOD,300 MHz) δ1.5-1.9(3H+CD3COOD,m);1.95(3H,s);2.05-2.35(2H,m);2.4-2.6(2H+DMSO,m);3.0-3.1(1H,m);3.2-3.9(4H,m);4.2-4.3(1H,m);6.5-6.7(2H,m);7.2-7.4(6H,m).

MS (ESP+) m/z 460 (M+H)⁺,311.

Anal.Calcd for C₂₃H₂₉N₃O₃S₂,1.35TFA C,50.3;H,4.99;N,6.85.

Found C,50.2;H,5.1;N,6.8.

EXAMPLE 40

Pharmaceutical Compositions

The following illustrate representative pharmaceutical dosage forms ofthe invention as defined herein (the active ingredient being termed“Compound X”), for therapeutic or prophylactic use in humans:

(a) Tablet I mg/tablet Compound X 100 Lactose Ph.Eur 182.75Croscarmellose sodium 12.0 Maize starch paste (5% w/v paste) 2.25Magnesium stearate 3.0 (b) Tablet II mg/tablet Compound X 50 LactosePh.Eur 223.75 Croscarmellose sodium 6.0 Maize starch 15.0Polyvinylpyrrolidone (5% w/v paste) 2.25 Magnesium stearate 3.0 (c)Tablet III mg/tablet Compound X 1.0 Lactose Ph.Eur 93.25 Croscarmellosesodium 4.0 Maize starch paste (5% w/v paste) 0.75 Magnesium stearate 1.0(d) Capsule mg/capsule Compound X 10 Lactose Ph.Eur 488.5 Magnesium 1.5(e) Injection I (50 mg/ml) Compound X 5.0% w/v 1 M Sodium hydroxidesolution 15.0% v/v 0.1 M Hydrochloric acid  (to adjust pH to 7.6)Polyethylene glycol 400 4.5% w/v Water for injection to 100% (f)Injection II (10 mg/ml) Compound X 1.0% w/v Sodium phosphate BP 3.6% w/v0.1 M Sodium hydroxide solution 15.0% v/v Water for injection to 100% (1mg/ml, buf-) (g) Injection III fered to pH 6 Compound X 0.1% w/v Sodiumphosphate BP 2.26% w/v Citric acid 0.38% w/v Polyethylene glycol 4003.5% w/v Water for injection to 100% (h) Aerosol I mg/ml Compound X 10.0Sorbitan trioleate 13.5 Trichlorofluoromethane 910.0Dichlorodifluoromethane 490.0 (i) Aerosol II mg/ml Compound X 0.2Sorbitan trioleate 0.27 Trichlorofluoromethane 70.0Dichlorodifluoromethane 280.0 Dichlorotetrafluoroethane 1094.0 (j)Aerosol III mg/ml Compound X 2.5 Sorbitan trioleate 3.38Trichlorofluoromethane 67.5 Dichlorodifluoromethane 1086.0Dichlorotetrafluoroethane 191.6 (k) Aerosol IV mg/ml Compound X 2.5 Soyalecithin 2.7 Trichlorofluoromethane 67.5 Dichlorodifluoromethane 1086.0Dichlorotetrafluoroethane 191.6 (l) Ointment ml Compound X 40 mg Ethanol300 μl Water 300 μl 1-Dodecylazacycloheptan-2-one 50 μl Propylene glycolto 1 ml Note The above formulations may be obtained by conventionalprocedures well known in the pharmaceutical art. The tablets (a)-(c) mayby enteric coated by conventional means, for example to provide a# coating of cellulose acetate phthalate. The aerosol formulations(h)-(k) may be used in conjunction with standard, metered dose aerosoldispensers, and the suspending agents sorbitan trioleate and soyalecithin may be replaced by an alternative suspending agent such assorbitan monooleate, sorbitan # sesquioleate, polysorbate 80,polyglycerol oleate or oleic acid.

EXAMPLE 41

(See Scheme 47)

Preparation of a)(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyricacid (compound 39e); and b)(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyricacid methyl ester (compound 39) a) Preparation of Compound 39

Compound 39a

32a (1.5 g, 6.2 mmol) was coupled with L-Glutamine methyl ester(analogously as for the equivalent step in Example 30) to give compound39a as a white solid, 1.2 g(50.5%)

MS (ESP)+ m/z 386 (M+H)⁺.

Compound 39

39a was reduced coupled with the aldehyde (22b) and selectivelydeprotected using the same methodology as previously described forExample 32 to give the title compound 39.

MS (ESP+) m/z 471 (M+H)⁺.

Anal.Calcd for C₂₄H₃N₄O₄S,3HCl,0.25H₂O C,49.3;H,5.8;N,9.6.

Found C,49.2;H,5.9;N,9.2.

b) Preparation of Compound 39e

39 was hydrolysed (analogously as for the equivalent step in Example 32)to give the title compound 39e.

MS (ESP−) m/z 455 (M−H)⁻.

Anal.Calcd for C₂₃H₂₈N₄O₄S,2TFA C,47.4;H,4.4;N,8.2.

Found C,47.0;H,4.5;N,7.9.

What is claimed is:
 1. A compound of any of the following classes i),ii) or iii): class i)

 wherein: X¹ is selected from H; C₁₋₆alkyl; hydroxyC₁₋₆alkyl,C₁₋₆alkoxyC₁₋₆alkyl; C₁₋₆alkylcarbonyl; hydroxyC₁₋₆alkylcarbonyl;C₁₋₆alkoxyC₁₋₆alkylcarbonyl; A is selected from phenyl and naphthyl; X²is selected from H; phenyl; phenylC₁₋₆alkyl; and a 5-6 memberedheteroaryl ring containing upto 3 heteroatoms selected from O, N and Soptionally linked to A by C₁₋₆alkyl; and X² is unsubstituted orsubstituted on any ring with substitutes selected from C₁₋₄alkyl,halogen, OH, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, amino, C₁₋₄alkylamino,di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, nitro, cyano, carboxy, carbamoyl,C₁₋₄alkoxycarbonyl, thiol, C₁₋₄alkylsulfanyl, C₁₋₄alkylsulfinyl,C₁₋₄alkylsulfonyl, and sulfonamido; X³ is selected from H; C₁₋₆alkyl; X⁴is selected from C₁₋₆alkylsulfanyl; C₁₋₆alkysulfinyl; C₁₋₆alkylsulfonyl;carbamoyl; N-(C₁₋₆alkyl)carbamoyl; N-(diC₁₋₆alkyl)carbamoyl; and hydroxyor a C₁₋₄alkyl ether thereof; class ii)

 wherein: X⁵ is selected from —CO—C₁₋₄alkyl-Ph: —CO—C₁₋₆alkyl;—CO—C₁₋₄alkyl-heteroaryl where heteroaryl is a 5-10 membered heteroarylring containing upto 5 heteroatoms selected from O, N and S and Ph orheteroaryl are unsubstituted or substituted with substituents selectedfrom C₁₋₄alkyl, halogen, OH, C₁₋₄alkanoyl, C₁₋₄alkylsulfinyl,C₁₋₄alkylsulfonyl and sulsonamido; C₁₋₄alkyloxyC₁₋₄alkyl; A is naphthylor a 10 membered heterocyclic ring having upto 5 heteroatoms selectedfrom O, N and S; R³ is selected from H; OH; CN; CF₃; NO₂;—C₁₋₄alkylene—R⁷ where R⁷ is selected from phenyl, naphthyl, and a 5-10membered monocyclic or bicyclic heteroaryl ring containing up to 5heteroatoms selected from O, N and S and any aryl ring in R⁷ isunsubstituted or substituted with substituents selected from C₁₋₄alkyl,halogen, OH, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, amino,C₁₋₄alkylamino, di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, nitro, cyano,carboxy, carbamoyl, C₁₋₄alkoxycarbonyl, thiol, C₁₋₄alkylsulfanyl,C₁₋₄alkylsulfinyl, C₁₋₄alkylsulfonyl and sulfonamido; R⁷; C₂₋₄alkenyl;halogen; —(CH₂)nCOOR⁸ where n=0-3 and R⁸ represents H, C₁₋₄alkyl,C₂₋₄alkenyl; —CONR⁹R¹⁰ where R⁹ R¹⁰ independently represent H,C₁₋₄alkyl, C₂₋₄alkenyl, —O—C₁₋₄alkyl, —O—C₂₋₄alkenyl, or —C₁₋₃alkylenePH which is unsubstituted or substituted with substituents selected fromC₁₋₄alkyl, halogen, OH, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy,amino, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, nitro,cyano, carboxy, carbamoyl, C₁₋₄alkoxycarbonyl, thiol, C₁₋₄alkylsulfanyl,C₁₋₄alkylsulfinyl, C₁₋₄alkylsulfonyl and sulfonamido; —CON(R¹¹)OR¹²where R¹¹ and R¹² independently represent H, C₁₋₄alkyl and C₂₋₄alkenyl;a group for Formula II, —CONR¹³—CHR¹⁴—COOR¹⁷, where R¹³ is H orC₁₋₄alkyl, R¹⁷ is H or C₁₋₆alkyl, R¹⁴ is selected from the side chain ofa lipophilin amino acid, carbamoyl C₁₋₄alkyl, N—(monoC₁₋₄alkyl)carbamoyl C₁₋₄alkyl and N—(di C₁₋₄alkyl)carbamoyl C₁₋₄alkyl,the group of Formula II having L or D configuration at the chiral alphacabon in the corresponding free amino acid; a lactone of formula

C₁₋₄alkyl monosubstituted on carbon with ═N—OH; a group of Formula—X—R¹⁵ where X is selected from O, CO, CH₂, S, SO, SO₂ and R¹⁶ isselected from C₁₋₆alkyl, phenyl, naphthyl, a 5-10 membered monocyclic orbicyclic heteroaryl ring containing up to 5 heteroatoms selected from O,N and S and any aryl ring in R¹⁵ is unsubstituted or substituted withsubstituents selected from C₁₋₄alkyl, halogen, OH, C₁₋₄alkoxy,C₁₋₄alkanoyl, C₁₋₄alkanoyloxy, amino, C₁₋₄alkylamino,di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, nitro, cyano, carboxy, carbamoyl,C₁₋₄alkoxycarbonyl, thiol, C₁₋₄alkylsulfanyl, C₁₋₄alkylsulfinyl,C₁₋₄alkylsulfonyl and sulfonamido; P is 0-3 which R³ values can be thesame or different; class iii)

 wherein: X⁶ has any value defined for X⁵ in ii) above; X⁷ isunsubstituted or substituted Ph with substituents selected fromC₁₋₄alkyl, halogen, OH, C₁₋₄alkoxy, C₁₋₄alkanoyl, C₁₋₄alkanoyloxy,amino, C₁₋₄alkylamino, di(C₁₋₄alkyl)amino, C₁₋₄alkanoylamino, nitro,cyano, carboxy, carbamoyl, C₁₋₄alkoxycarbonyl, thiol, C₁₋₄alkylsulfanyl,C₁₋₄alkylsulfinyl, C₁₋₄alkylsulfonyl and sulfonamido; A is Ph ornaphthyl or a 5-10 membered heterocyclic ring having upto 5 heteroatomsselected from O, N and S; R³ and p are as defined in ii) above; or aN-oxide, pharmaceutically acceptable salt, prodrug or solvate thereof.2. A compound according to claim 1 which: in compounds of class i), X¹is selected from H and C₁₋₆alkoxyC₁₋₆alkyl; X² is selected from H:phenyl or phenylC₁₋₆alkyl; X⁴ is C₁₋₆alkylsulfanyl; A is selected fromphenyl or naphthyl; in compounds of class ii), p is 0 and; in compoundsof class iii) X⁷ is Ph; A is Ph; p is
 0. 3. Any one of the followingcompounds or a pharmaceutically acceptable salt thereof:(2S)-2-{2-Benzyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid methyl ester;(2S)-2-{2-Benzyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-benzoylamino}-4-methylsulfanylbutyricacid;(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester;(2S)-2-({2-phenyl-5-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid;(2S)-2-({3-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester;(2S)-2-({3-[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-naphthalene-1-carbonyl}-amino)-4-methylsulfanylbutyricacid;(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid methyl ester;(2S)-2-({-3-phenyl-5[([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-4-methylsulfanylbutyricacid;(2S,4S)-2-[{N-(4-methoxybenzyl)-N-(naphthalen-1-ylmethyl)-amino}-methyl]-pyrrolidine-4-thiol;N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-pentanamide;N-(naphthalen-1-ylmethyl)-N-([2S,4S]-4-sulfanylpyrrolidin-2-ylmethyl)-2-(pyridin-3-yl)-acetamide;N-((2S,4S)-4-sulfanyl-pyrrolidin-2-ylmethyl)-3-methyl-N-(2-naphthalen-1-yl-ethyl)butyramide;N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-N-(2-naphthalen-1-yl-ethyl)-2-pyridin-3-yl-acetamide;(2S,4S)-2-{[(3-Methoxypropyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol;N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-2-(4-methoxy-phenyl)-N-(2-naphthalen-2-yl-ethyl)-acetamide;(2S,4S)-2-{[(2-(4-Methoxyphenyl)ethyl)-(2-naphthalen-1-ylethyl)amino]methyl}-pyrrolidine-4-thiol;N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidine-2-ylmethyl)-3-methyl-butyramide;N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-2-yl-ethyl)-butyramide;N-(2,2-Diphenyl-ethyl)-N-([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-butyramide;(2S)-2-{3-[([2S,4S]-4-Sulfanyl-pyrrolidin-2-ylmethyl)-(3-methoxy-propyl)-amino]-benzoylamino}-4-methylsulfanyl-butyricacid;N-([2S,4S]-4-Sulfanyl-pyrrolidin-2-ylmethyl)-3,3-dimethyl-N-(2-naphthalen-1-yl-ethyl)-butyramide;(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyricacid; or(2S)-4-Carbamoyl-2-({2-phenyl-5-[([2S,4S]-4-sulfanyl-pyrrolidin-2-ylmethyl)-amino]-phenylcarbonyl}-amino)-butyricacid methyl ester.
 4. A pharmaceutical composition comprising a compoundas defined in any one of claims 1-3 together with a pharmaceuticallyacceptable diluent or carrier.
 5. A pharmaceutical composition accordingto claim 4 which is in the form of a tablet.
 6. A method for thetreatment of disease mediated through farnesylation of ras whichcomprises the administration of a pharmaceutical composition as claimedin claims 1, 2, or
 3. 7. A method for the treatment of a diseasemediated through farnesylation of ras which comprises the administrationof a pharmaceutical composition as claimed in claim
 4. 8. A compound ofthe Formula VII:

 wherein: R² is selected from H and —COC₁₋₄alkyl; X¹ is H; X² isselected from phenyl and phenylC₁₋₆alkyl; X³ is selected from H orC₁₋₆alkyl; X⁴ is C₁₋₆alkylsulfanyl; and A is phenyl.
 9. A methodaccording to claim 1 of treating carcinoma comprising administration ofan effective amount of a compound of Formula III or a pharmaceuticallyacceptable salt or prodrug thereof to a mammal in need of suchtreatment.
 10. A method according to claim 9 of treating a carcinomaselected from bladder, breast, colon, kidney, liver, lung, ovary,pancreas, stomach, cervix, thyroid, and skin.
 11. A method according toclaim 1 of treating hematopoietic tumors of lymphoid lineage comprisingadministering to a mammal requiring such treatment an effective amountof a compound of Formula III or a pharmaceutically acceptable salt orprodrug thererof.